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The number of described species of Brachycephalus has rapidly increased in the last decade (n = 22, which represents 56% of the total). Species of the genus Brachycephalus are mostly distributed in isolated mountaintops from Bahia (northeastern Brazil) to Santa Catarina states (southern Brazil), each one occupying only one or a few adjacent mountaintops. Herein, we described a new species of Brachycephalus of the B. pernix group, from Serra do Tabuleiro in Santa Catarina state, which also represents the southernmost known species. The new species can be distinguished from its congeners by a combination of characters, including the following: (1) "bufoniform" body; (2) small adult SVL: 9.57–11.10 mm for males and 10.88–12.70 mm for females; (3) head proportionally small (HL/SVL 19–28%) and eye proportionally large (ED/HL 36–56%); (4) dorsum texture rough; (5) snout shape rounded in dorsal and lateral views; (6) general dorsal body color olive green with head, arms and legs yellow-orangish scattered with olive green, and an orangish vertebral stripe spotted with white and brown colors; (7) skull and skeleton without hyperossification; (8) frontoparietal and sphenethmoid not fused; (9) advertisement with one or two high-frequency notes (6,115–6,562 Hz), and 2–4 pulses per note. The type locality is adjacent to Parque Estadual da Serra do Tabuleiro, a protected area, but we observed various agricultural activities in this locality, including the presence of exotic plants, which can change the amount and the quality of leaf litter, somehow compromising the population of the new species. Another aggravating factor is that the municipality of São Bonifácio has conflicts over land use with irregular occupation and unfinished expropriation processes in Parque Estadual da Serra do Tabuleiro. Considering that Brachycephalus sp. nov. is probably a mountaintop microendemic species, it is paramount that future studies quantifying the new species' full distribution and evaluating population trends to accurately assess its conservation status.

Made available in DSpace on 2022-04-28T19:41:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-30 Fundação Grupo Boticário de Proteção à Natureza Universidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências, Praça Infante Dom Henrique s/n Mater Natura - Instituto de Estudos Ambientais, Rua Emiliano Perneta, 297, sala 122 Universidade Federal Do Paraná Departamento de Zoologia Universidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências, Praça Infante Dom Henrique s/n

Made available in DSpace on 2022-04-29T08:35:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) The genus Brachycephalus includes small species of aposematic anurans known as microendemic, occurring in the mountains of the Atlantic Forest. Brachycephalus ephippium, B. nodoterga and B. pernix have been reported to contain the neurotoxin tetrodotoxin in skin and viscera. The biological conservation of several Brachycephalus species is currently threatened by climate change, deforestation, and the pandemic caused by the fungus Batrachochytrium dendrobatidis (Bd). Despite the well-known importance of amphibians’ associated bacteria in the defensive role against pathogens, there is still a poor understanding of amphibian microbiome composition. The present study investigated the composition of B. pitanga microbial community and the presence of TTX in the host and in cultures of bacterial isolates, using a combination of metagenomics, bacterial culture isolation, mass spectrometry and metabolomic analyses. Results of culture-dependent and -independent analyses characterized the microbial communities associated with the skin and viscera of B. pitanga. Mass spectrometry analysis indicated the presence of TTX in host tissues, while bacterial production of TTX was not observed under the experimental conditions used in this investigation. This is the first report confirming the occurrence of TTX in B. pitanga. Instituto de Química de São Carlos Universidade de São Paulo, CP 780 Instituto de Biologia SAGE-COPPE Universidade Federal do Rio de Janeiro (UFRJ) Departamento de Tecnologia Bioquímico-Farmacêutica (FBT) Faculdade de Ciências Farmacêuticas Universidade de São Paulo Laboratório de Toxinologia Instituto de Biologia Universidade de Brasília Departamento de Biodiversidade e Centro de Aquicultura Instituto de Biociências Universidade Estadual Paulista (UNESP) Instituto D'Or de Pesquisa e Ensino (IDOR) Instituto de Ciências Biomédicas Universidade Federal do Rio de Janeiro (UFRJ) Departamento de Biodiversidade e Centro de Aquicultura Instituto de Biociências Universidade Estadual Paulista (UNESP) FAPESP: 2013/50228-8 FAPESP: 2013/50741-7 FAPESP: 2016/14375-4 FAPESP: 2016/21341-9 FAPESP: 2019/00447-1 FAPERJ: 202.488/2019

Made available in DSpace on 2022-04-29T08:29:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Four species of pumpkin toadlets are currently recognized by the extreme condition of hyperossification in the skull and vertebral column within the genus Brachycephalus. In addition to their larger body size, relative to other congeneric species, Brachycephalus darkside, Brachycephalus ephippium, Brachycephalus garbeanus, and Brachycephalus margaritatus share remarkable osteological features, such as the large paravertebral plates forming an ornamented dorsal bone shield, visible through the integument. We add to the current knowledge of this group by analyzing its diversity and describing a new hyperossified species from some important Atlantic Forest remnants in southeastern Brazil, including those bordering São Paulo, the largest and most populous city in South America. The new species is diagnosed by a combination of morphological, osteological, and bioacoustic characters, with further evidence of mtDNA sequences to confirm its distinction from the other congeners. The parotic and paravertebral bone plates externally bordered with a rough and pale contour can readily distinguish the new species from the other hyperossified species within Brachycephalus. The new species is widely distributed along the Serra do Mar mountain range, including the regionally named Serra de Paranapiacaba, in elevations from 700 to 1000 m above sea level (a.s.l.), in the central coast of the state of São Paulo, southeastern Brazil. We discuss valuable morphological characters within this group of species, reinforce the need for further studies, and highlight the relevance of protected areas for biodiversity conservation in metropolitan regions. Instituto Nacional da Mata Atlântica (INMA) Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP) Universidade Estadual Paulista (UNESP) Instituto de Biociências Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP) Fundacao Para A Conservacao e A Producao Florestal Do Estado de Sao Paulo Parque Estadual da Serra Do Mar Núcleo São Sebastião Laboratório de Coleções Zoológicas Instituto Butantan Comitê Brasileiro de Registros Ornitológicos Www.cbro.org.br Universidade Federal de Minas Gerais Instituto de Ciências Biológicas Laboratório de Herpetologia Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP) Universidade Estadual Paulista (UNESP) Instituto de Biociências Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP)

Background. Brachycephalus includes miniaturized frogs with restricted geographical distributions throughout the Brazilian Atlantic Forest. Ecological data for most species are still scarce. For instance, advertisement calls have only been described for 12 of the 36 known species, including B. tridactylus, a recently described species from southern Brazil. Posteriorly, features of the advertisement call of B. tridactylus were compared with congeners and the unique characteristics of its call were highlighted. To confirm these potentially divergent characteristics, we reanalysed an original recording of B. tridactylus and analysed our own recordings and verified that the original description of its advertisement call is inaccurate. Thus, we redescribe its advertisement calls. Methods. We asked the descriptors of B. tridactylus the original recordings that they made and requested access to the only original recording deposited in a collection of sounds. We received from André Lima a copy of one recording, the same as the one that had deposited, and obtained permission to re-analyze it. We studied this recording and compared it with our own recordings, made at the type locality of the species on March, 2016. Sound samples were analysed with Raven Pro 1.5.0 and call analyses were made under a note-centered approach. Results. The original recording was amplified somehow by at least 6 dB and was also clearly low-pass filtered with a cutoff frequency of 10 kHz. Our analyses did not allow us to recognize several of the acoustic parameters normally described in Brachycephalus. The sound we heard from the notes overlapped with other signals (noise?), which prevented us from clearly determining the end of the note and other important features, such as the presence of pulses. According to our recordings (n = 15 individuals), B. tridactylus emitted a relatively long advertisement call (50.8 s, on average), composed by 10–13 notes emitted in a note rate of 3.7–8.3 notes per minute. Only isolated notes were present. The notes were composed by 1–3 pulses and the note duration varied from 0.002–0.021 s. Discussion. The original description of the call of B. tridactylus is incorrect because it included background noise and amplification artefacts as part of the call parameters. However, we recognize that the original recording and our recordings have captured the same type of call. In our measurements of the species calls, note duration was nearly an order of magnitude shorter as the original description. The existence of notes with 1–3 pulses was not acknowledged in the original description. With few pulses per notes, the advertisement call of B. tridactylus is distinct from the notes with several pulses of B. ephippium, B. pitanga, B. crispus, B. sulfuratus, and B. darkside. The advertisement calls of B. tridactylus is also distinct from that of B. albolineatus and B. mirissimus by having only isolates notes, instead of isolated notes and note groups.

Although we celebrate the centennial of Brachycephalus garbeanus’ discovery, little progress has been done on understanding this species’ biology apart from a few morphological and ecological studies, which includes its redescription based on three specimens from the type-series, microhabitat use, sexual dimorphism in body size, and feeding habits. This species is endemic to the Serra dos Órgãos Mountain range, in the state of Rio de Janeiro, southeastern Brazil. Here we redescribe B. garbeanus based on a wide sampling, including its advertisement and aggressive calls, and also the chigger mites infestation pattern. The advertisement call is longer than 25.8 s with pulsed notes series emitted at an average rate of 2.3 notes/s and 14.1 pulses/s; long inter-note interval with 320 ms; notes with distinctly short pulses (1 to 16 ms); low dominant frequency for this genus (3.0-5.4 kHz) and presence of four harmonics. This species is often parasitized by chigger mites of Hannemania, with a prevalence of infection of 67%, mainly affecting the ventral body surface. Females had a higher prevalence of parasites than males and there was no correlation found between the size of specimens and the number of parasites. Our study, provides an important and overdue taxonomical contribution, including a large amount of novel information for B. garbeanus.

We describe a new species of dull-colored flea-toad, genus Brachycephalus, from the Atlantic Forest of Caparaó mountains in southeastern Brazil. The new species is characterized by its diminutive size, “leptodactyliform” body, brownish color with an inverted V-shaped dark mark on dorsum, skin smooth, hyperossification and dorsal shield absent, linea masculinea absent, Fingers I and IV vestigial, Toe I externally absent, Toe II reduced but functional, Toes III and IV with pointed tips, Toe V vestigial, and ventral color uniformly brown. It is a leaf litter dweller, known only from type locality in the humid forests on the eastern slopes of Parque Nacional do Caparaó mountains, a protected area in the states of Espírito Santo and Minas Gerais, southeastern Brazil. It is the third flea-toad occurring in the state of Espírito Santo recovered as sister to all other Brachycephalus distributed from the state of São Paulo northward in the Atlantic Forest.

Brachycephalus bufonoides Miranda-Ribeiro, 1920 Brachycephalus ephippium var. bufonoides Miranda-Ribeiro, 1920. Brachycephalus ephippium: Cochran, 1955 (part). Brachycephalus bufonoides: Pombal, 2010. Lectotype. MZUSP 1459 (designated by Pombal 2010). Type locality: Serra de Maca��, Nova Friburgo, State of Rio de Janeiro, Brazil on September 1909 collected by Ernest Garbe. Paralectotype MZUSP 1458, collected with the lectotype. Referred specimens. The Serra Queimada trail (22��19���21������S, 42��16���19������W: datum WAG84; 1169 m above sea level), ��rea de Prote����o Ambiental (APA) de Maca�� de Cima, municipality of Nova Friburgo, state of Rio de Janeiro, souTHeasTern Brazil: MNRJ 91685 (male) anD ZUFRJ 15424���26 (males), 15428���30 (males) collecTeD on 3 OcTober 2015 by M. Folly anD L. C. Amaral; anD MNRJ 91686 (female), MNRJ 91687���88 (males), ZUFRJ 15525 (male), ZUFRJ 15527 (male), ZUFRJ 15528���29 (females), ZUFRJ 15530���31 (males), ZUFRJ 15534���35 (males), and ZUFRJ 15536 (female) collected on 20 November 2015 by Manuella Folly, Lucas C. Amaral, Sergio P. de Carvalho-e-Silva, and Jos�� P. Pombal Jr. Diagnosis. The specimens show absence of sternum; eight presacral vertebrae; palatal shelf of maxilla lacking pterygoid process; neopalatines and columella absents; arrow-shaped terminal phalanges; one phalange in Finger V, very short phalange in Toe I. These characters have been proposed as diagnostic of Brachycephalus (Hedges et al. 2008). Brachycephalus bufonoides belongs to this genus by the aforementioned characters and also by the small size (B. ephippium species group by: (1) bufoniform body shape; and (2) hyperossified skull and presence of hyperossified spinal processes of sacral and presacral vertebrae (intermediate condition sensu Clemente-Carvalho et al. 2012). Adults of B. bufonoides are distinguished from all of its congeners by the combination of the following characters: (1) skin on head and dorsum with dermal hyperossification; (2) skull with hyperossification of postorbital crests; (3) a pair of hyperossified bulges about equidistant between postorbital crests; (4) fourth presacral vertebra with transverse process hyperossified, ornamented and sacral diapophyses hyperossified, which can be seen externally (lineage of B. vertebralis sensu Condez et al. 2020); (5) presence of dermal ossification as separated bulges of each vertebrae; (6) general background color orange with different intensities of dark orange blotches on dorsum, including bordering of sacral region; (7) absence of osteoderms and presence of warts on the dorsolateral surface of body; (8) medium body size (SVL of adults: 12.0��� 14.5 mm for males and 14.7���16.3 mm for females; Table 1); (9) rough dorsum; (10) advertisement calls with 13 to 17 pulses; (11) presence of pulse period modulation; and (12) advertisement calls with notes longer than 0.2 s (0.22 to 0.31 s). Comparisons with the other species. Brachycephalus bufonoides (Fig. 2) has skin on the head and vertebrae with dermal ossification, and a hyperossified process of the fourth vertebra. These characteristics distinguish the species from B. pernix species group (B. actaeus, B. albolineatus, B. auroguttatus, B. boticario, B. brunneus, B. coloratus, B. curupira, B. ferruginus, B. fuscolineatus, B. izecksohni, B. leopardus, B. mariaeterezae, B. mirissimus, B. olivaceus, B. pernix, B. pombali, B. quiririensis, B. tridactylus, and B. verrucosus; Pombal et al. 1998; Ribeiro et al. 2005; Alves et al. 2006; Haddad et al. 2010; Garey et al. 2012; Pie and Ribeiro 2015; Bornschein et al. 2016; Monteiro et al. 2018a; Ribeiro et al. 2017; Pie et al. 2018) and from B. didactylus, B. hermogenesi, B. pulex, and B. sulfuratus that completely lack hyperossification (Izecksohn, 1971; Giaretta and Sawaya, 1998; Napoli et al. 2011; Clemente-Carvalho et al. 2012; Condez et al. 2016). By having a hyperossified dorsal shield or hyperossified spinal process of sacral and presacral vertebrae, Brachycephalus bufonoides belongs to B. ephippium species group. This species can be distinguished from B. darkside, B. ephippium, B. garbeanus, and B. margaritatus by its presence of dermal ossification on the vertebrae (bony shields on dorsum in these species: Pombal 2010; Pombal and Izecksohn 2011; Guimar��es et al. 2017). Brachycephalus bufonoides has a transverse process of the fourth presacral vertebra and sacral diapophyses hyperossified and ornamented, which is very distinct of the large bone plate exhibited by B. ephippium, B. darkside, B. garbeanus, and B. margaritatus; and also distinct from the comparatively less ornamented skeleton (intermediate condition sensu Clemente-Carvalho et al. 2012; B. vertebralis lineage sensu Condez et al. 2020); B. vertebralis lineage sensu Condez et al. 2020 of B. alipioi, B. crispus, B. guarani, B. nodoterga, B. pitanga, B. toby, and B. vertebralis (Pombal and Gasparini 2006; Haddad et al. 2010; Clemente-Carvalho et al. 2012; Condez et al. 2014). Absence of osteoderms distinguishes B. bufonoides species from B. crispus, B. margaritatus, and B. nodoterga. Brachycephalus bufonoides has skin on its head with dermal ossification and the presence of dermal ossification on the vertebrae while B. atelopoide has no dorsal shields and cephalic ossification (Miranda-Ribeiro 1920; Pombal 2010).The orange coloration with different intensities of dark orange blotches on the dorsum of B. bufonoides differs from B. alipioi, B. pitanga, B. guarani, and B. toby (orange uniform in B. alipioi: Pombal and Gasparini 2006); orange with red irregular blotches in B. pitanga (Alves et al. 2009); orange with a dark brown vertebrate stripe in B. guarani (Clemente-Carvalho et al. 2012); orange with dorsal greenish in B. toby (Haddad et al. 2010). Further, B. bufonoides can be distinguished from B. crispus and B. nodoterga by its dorsum without dermal ossification like warts on the skin [with ossification is B. crispus and B. nodoterga (Condez et al. 2014)]. Redescription of the species. Measurements for 15 males and four females are given in Table 1. Vocal slits not observed in males; body robust and bufoniform (Figs. 1���3); head wider than long (HL/HW x = 0.51 in males and females); head length approximately 19% of SVL in males and 20% in females; snout short with length almost half of eye diameter (END/ED x = 0.56 in males and 0.57 in females), rounded in dorsal and lateral views; nostrils, directed anterolaterally, elliptical, not protuberant; canthus rostralis distinct, almost straight; loreal region vertical; eye not protruding dorsally, mean of eye diameter 57% of HL in males and 46% in females; tympanum not visible; supratympanic fold absent; a hyperossified postorbital crest; a pair of hyperossified bulges about equidistant between postorbital crest; tongue long and narrow, without indentation on its free posterior border; choanae elliptical, anterior to the eye; vomerine odontophores absent; premaxillary and maxillary teeth absent. Arm slightly shorter than forearm (AL/FAL x = 0.90 in males and 0.87 in females), slender; total arm length with mean of 42% SVL in males and 39% in females; hands with the same length in males (AL/HAL x = 1.0) and almost of the same length of upper arm in females (AL/HAL x = 0.95); Finger III and IV robust, distinct; IV longer than III; Finger II very short and Finger V vestigial; pointed tip of Finger III and IV; subarticular tubercles, inner, and outer metacarpal tubercles absent. Shank as long as thigh (SL/TL x = 0.96 in males and 0.95 in females); total leg length with mean of 84% of SVL in both males and females; thigh length larger than foot length (TL/FL x = 0.78 in males and 0.80 in females); Toe II, III, and IV distinct, robust; Toe I and V reduced; tip of Toe II rounded, tip of Toe III and IV pointed; relative lengTH of Toes I ��� V Coloration in life. General background coloration orange; dorsum with a dark orange blotch bordering the spine; protruding dorsal hyperossifications lighter orange than the dorsum color (Fig. 1); finger IV and toe V with black tips; a light-yellow line below the eye, eye black in color. Dorsum varies from orange to light orange blotches bordering to the spine varying in different intensities. Coloration in preservative and variation. Upper surfaces gray (Figs. 3 A���D); protruding dorsal hyperossifications beige (Fig. 3E), flanks light brown; tips of Finger IV and toe V gray; a cream line below the eyes. Dorsal surfaces of body can be gray to brown, varying specially from below the head to the cloaca. In robust specimens, the ossified structures on the head, dorsum and diapophyses are less distinct (Fig. 3D) than thinner specimens that have such ossified structures much evident (Figs. 3A, C). The hyperossification in the spinal processes of presacral vertebrae is completely ossified forming an irregular (MNRJ 91688 and MNRJ 91687) or regular oval shape (ZU-FRJ 15428 and ZUFRJ 15529). Natural history. Specimens of Brachycephalus bufonoides were active during the day were found on forest floor or amidst leaf litter. Some individuals were found under fallen tree trunks where the moisture is higher. Specimens were observed walking slowly on the leaf-litter. Generally, males were observed and heard calling under the leaves of the leaf-litter or branches, but they can be found calling exposed on the surface of leaf-litter or timber (males observed on cloudy and rainy days). Individuals were seen calling from both above and under the leaf-litter, except for one individual, which was calling on a fallen tree trunk. Vocalizations. The individuals found calling in the field were recorded at air temperature of 20.8 oC, and relative humidity of the air of 78%. Calls were emitted as a regular series of low-intensity buzzes (around 120 dB at about 1-meter recording distance). The advertisement call duration is of up to 3 minutes (specimens usually stopped calling when disturbed), and consists of a series of regularly emitted notes, with constant general amplitude (Fig. 4). Notes consisted of a number of pulses, ranging from 13 to 17 pulses/note (x = 15.05 �� 0.88; = 15; Mo = 16; n = 352), with most of them consisting of 14 to 16 pulses (96.7%). Amplitude modulation increased until the second quarter, with a mild descending modulation from end of second quarter until the end of the note. Note duration ranges from 222.09 to 308.53 ms (x = 271.33 �� 18.70; = 276.00; Mo = 290.07; n = 352). Inter-note interval duration ranges from 147.58 to 265.00 ms (x = 209.05 �� 24.68; = 219.00; Mo = 232.00; n = 298). Note period ranges from 401.82 to 545.00 ms (x = 482.10 �� 35.61; = 495.00; Mo = 503.00; n = 298). Note repetition rate ranges from 1.98 to 2.43 note/s (x = 2.15 �� 0.20; = 2.04; n = 3). Pulse period ranges from 9.89 to 25.13 ms (x = 18.18 �� 3.39; n = 729). There is a pulse period modulation to shorter duration of the periods starting through the second third of the note, and continuing modulation to longer duration of the periods through the final third of the note. Pulse repetition rate ranges from 47.55 to 71.28 pulses/s (x = 56.20 �� 4.81; = 55.01; Mo = 51.27; n = 352). Pulse envelope with decay steeper than attack. Dominant frequency of the call ranges from 4.13 to 4.88 kHz (x = 4.55 �� 0.14; = 4.5; Mo = 4.5; n = 344); minimum frequency ranges from 2.49 to 4.24 kHz (x = 3.38 �� 0.37; = 3.43; Mo = 3.37; n = 98), and maximum frequency ranges from 5.58 to 7.05 kHz (x = 6.268 �� 0.37; = 6.25; Mo = 5.94; n = 98). Comparison with advertisement calls of species of Brachycephalus. Brachycephalus bufonoides differs from the other species of Brachycephalus by a number of distinct advertisement call parameters. Brachycephalus bufonoides has greater number of pulses (range = 13���17) than the species belonging to the B. pernix group: B. actaeus 2���3 pulses (Monteiro et al. 2018a); B. albolineatus 1���3 pulses (Bornschein et al. 2017); B. mirissimus 1���3 pulses (Pie et al. 2018); B. olivaceus 2���3 (Monteiro et al. 2018b); B. pernix: 3 pulses (Wistuba 1998); B. quiririensis 3���4 pulses (Monteiro et al. 2018b); B. tridactylus 1 pulse (Garey et al. 2012), and also presents longer note duration in B. bufonoides: x = 271.33 ms, while in B. actaeus x = 4 ms (Monteiro et al. 2018a); B. albolineatus x = 20 ms (Bornschein et al. 2017); B. olivaceus x = 38 ms (Monteiro et al. 2018b); B. pernix x = 60 ms (Wistuba 1998); B. quiririensis x = 50 ms (Monteiro et al. 2018b); B. tridactylus x = 110 ms (Garey et al. 2012). It also differs acoustically from most of them by presenting shorter inter-note interval in B. bufonoides x = 0.27 s, while in B. actaeus x = 5.42 s (Monteiro et al. 2018a); B. albolineatus x = 6.66 s (Bornschein et al. 2018); B. mirissimus x = 5.83 s (Pie et al. 2018); B. olivaceus: x = 10.69 s (Monteiro et al. 2018b); B. quiririensis: x = 12.58 s (Monteiro et al. 2018b), and lower dominant frequency in B. bufonoides x = 4.6 kHz, while in B. actaeus x = 6.9 kHz (Monteiro et al. 2018a); B. albolineatus x = 6.4 kHz (Bornschein et al. 2018); B. mirissimus x = 6.7 kHz (Pie et al. 2018); B. olivaceus: x = 6.8 kHz (Monteiro et al. 2018b); B. quiririensis x = 6.3 kHz (Monteiro et al. 2018b). The dominant frequency of B. tridactylus x = 4.8 kHz (Garey et al. 2012) overlaps with the range of B. bufonoides (4.1���4.9 kHz), and so does B. pernix 4.5���6.7, x = 5.6 kHz (Wistuba 1998). Values for pulse repetition rate and note repetition rate also vary greatly. The species from Brachycephalus ephippium group have an overall similar structure of the call, consisting in constant repetition of pulsed notes. Brachycephalus bufonoides can be distinguished from the other species of the B. ephippium group by a combination of different call structures. If differs most from B. darkside for presenting greater number of pulses (B. bufonoides 13���17 pulses, x = 15) in B. darkside 5���8 pulses, x = 6 (Guimar��es et al. 2017), while is similar or overlaps parts of the range with the other species: B. crispus 7���12 pulses, x = 10 (Condez et al. 2014); B. ephippium 5���15 pulses, x = 12 (Pombal et al. 1994); B. pitanga: 7���14 pulses, x = 11 (Ara��jo et al. 2012; Tandel et al. 2014). The note duration also is close to or overlaps some of its range with two of the species (B. bufonoides: 221���309, x = 271 ms): B. crispus x = 280 ms (Condez et al. 2014); B. pitanga: 150���250, x = 190 ms (Tandel et al. 2014), x = 170 ms (Ara��jo et al. 2012), but the value is lower in B. darkside x = 111 ms: Guimar��es et al. 2017) and B. ephippium (x = 112 ms: Pombal et al. 1994). The interval between notes of B. bufonoides (148���265, x = 209 ms) differs mostly from that of B. crispus (x = 350 ms: Condez et al. 2014), but overlaps slightly with the other species (B. darkside 122���215, x = 159.5 ms: Guimar��es et al. 2017); B. ephippium (123���149, x = 134 ms: Pombal et al. 1994); B. pitanga (200���430, x = 280 ms: Tandel et al. 2014). The pulse repetition rate (x = 56.20 pulses/s in B. bufonoides) is distinct from that of B. crispus x = 17.4 pulses/s: Condez et al. 2014) but is similar to the rate of B. darkside x = 56.9 pulses/s: Guimar��es et al. 2017) and B. pitanga x = 62.4 pulses/s: Ara��jo et al. 2012). Furthermore, note repetition rate of B. bufonoides (x = 2.15 notes/s, or 129 notes/min) is similar to the rate of B. crispus x = 1.67 notes/s: Condez et al. 2014) and B. pitanga x = 159 notes/min: Ara��jo et al. 2012) but differs from that of B. darkside x = 211 notes/min: Guimar��es et al. 2017). Brachycephalus bufonoides can be further distinguished from B. darkside for its higher dominant frequency (B. bufonoides: 4.1���4.9 kHz, x = 4.6 kHz) while in B. darkside: 2.8���3.8 kHz (Guimar��es et al. 2017), but differ less from the other species (B. ephippium: x = 3.9 kHz (Goutte et al. 2017), lowest frequency = 3.4 kHz, highest frequency = 5.3 kHz (Pombal et al. 1994), and overlaps with the range of B. crispus: x = 4.6 kHz: Condez et al. 2014 and B. pitanga: x = 4.9 kHz (Ara��jo et al. 2012) and x = 4.8 kHz (Tandel et al. 2014). The advertisement call of B. hermogenesi (x = 6.8 kHz: Verdade et al. 2008) and B. sulfuratus (x = 6.7 kHz: Condez et al. 2016) differ from B. bufonoides by presenting higher dominant frequency. Some overlap between the note duration of B. sulfuratus (131���233, x = 195 ms: Condez et al. 2016) and B. bufonoides (221���309 ms) was noticed, however a greater number of pulses per note in B. bufonoides (x = 15 pulses) than in B. sulfuratus 7���11 pulses, x = 9 (Condez et al. 2016) was observed, which results in difference in the pulse rate between B. bufonoides (x = 56.2 pulses/s) and B. sulfuratus (6.1���12.3, x = 9.3 pulses/s: Condez et al. 2016). The note repetition rate of B. bufonoides (x = 2.15 notes/s) is greater than B. hermogenesi (, Published as part of Folly, Manuella, Amaral, Lucas Coutinho, Carvalho-E-Silva, Sergio Potsch De & Pombal Jr, Jos�� P., 2020, Rediscovery of the toadlet Brachycephalus bufonoides Miranda-Ribeiro, 1920 (Anura: Brachycephalidae) with osteological and acoustic descriptions, pp. 265-294 in Zootaxa 4819 (2) on pages 269-283, DOI: 10.11646/zootaxa.4819.2.3, http://zenodo.org/record/4396900, {"references":["Miranda-Ribeiro, A. (1920) Os Brachycephalideos do Museu Paulista (com tres especes novas). Revista do Museu Paulista, 12, 307 - 316. https: // doi. org / 10.5962 / bhl. part. 22255","Cochran, D. M. (1955) Frogs of southeastern Brazil. Bulletin of the United States National Museum, 206, 1 - 423. https: // doi. org / 10.5479 / si. 03629236.206.1","Pombal, J. P. Jr. (2010) A posicao taxonomica das variedades de Brachycephalus ephippium (Spix, 1824) descritas por Miranda- Ribeiro, 1920 (Amphibia, Anura, Brachycephalidae). Boletim do Museu Nacional (N. S.), Zoologia, 526, 1 - 12.","Hedges, S. B., Duellman, W. E. & Heinicke, M. P. (2008) New World direct-development frogs (Anura: Terrarana): Molecular phylogeny, classification, biogeography, and conservation. Zootaxa, 1737 (1), 1 - 182. https: // doi. org / 10.11646 / zootaxa. 1737.1.1","Clemente-Carvalho, R. B. G., Giaretta, A. A., Condez, T. H., Haddad, C. F. B. & dos Reis, S. F. (2012) A new species of miniaturized toadlet, genus Brachycephalus (Anura: Brachycephalidae), from the Atlantic forest of Southeastern Brazil. Herpetologica, 68 (3), 365 - 374. https: // doi. org / 10.1655 / HERPETOLOGICA-D- 11 - 00085.1","Condez, T. H., Haddad, C. F. B. & Zamudio, K. R. (2020) Historical biogeography and multi-trait evolution in miniature toadlets of the genus Brachycephalus (Anura: Brachycephalidae), Biological Journal of the Linnean Society, 129 (3), 664 - 686. https: // doi. org / 10.1093 / biolinnean / blz 200","Pombal, J. P. Jr., Wistuba, E. M. & Bornschein, M. R. (1998) A new species of brachycephalid (Anura) from the Atlantic Rain Forest of Brazil. Journal of Herpetology, 32 (1), 70 - 74. https: // doi. org / 10.2307 / 1565481","Ribeiro, L. F., Alves, A. C. R., Haddad, C. F. B. & Reis, S. F. dos (2005) Two new species of Brachycephalus Gunther, 1858 from the State of Parana, southern Brazil (Amphibia, Anura, Brachyephalidae). Boletim do Museu Nacional, New Series, Zoologia, 519, 1 - 18.","Alves, A. C. R., Ribeiro, L. F., Haddad, C. F. B. & Reis, S. F. (2006) Two new species of Brachycephalus (Anura: Brachycephalidae) from the Atlantic forest in Parana State, southern Brazil. Herpetologica, 62 (2), 221 - 233. https: // doi. org / 10.1655 / 05 - 41.1","Haddad, C. F. B., Alves, A. C. R., Clemente-Carvalho, R. B. G. & dos Reis, S. F. (2010) A new species of Brachycephalus from the Atlantic Rain Forest in Sao Paulo State, southeastern Brazil (Amphibia: Anura: Brachycephalidae). Copeia, 2010 (3), 410 - 420. https: // doi. org / 10.1643 / CH- 09 - 102","Garey, M. V., Lima, A. M. X., Hartmann, M. T. & Haddad, C. F. B. (2012) A new species of miniaturized toadlet, genus Brachy- cephalus (Anura: Brachycephalidae), from southern Brazil. Herpetologica, 68 (2), 266 - 271. https: // doi. org / 10.1655 / HERPETOLOGICA-D- 11 - 00074.1","Ribeiro, L. F., Bornschein, M. R., Belmonte-Lopes, R., Firkowski, C. R., Morato, S. A. A. & Pie, M. R. (2015) Seven new microendemic species of Brachycephalus (Anura: Brachycephalidae) from southern Brazil. PeerJ, 3, e 1011. https: // doi. org / 10.7717 / peerj. 1011","Bornschein, M. R., Ribeiro, L. F., Blackburn, D. C., Stanley, E. L. & Pie, M. R. (2016) A new species of Brachycephalus (Anura: Brachycephalidae) from Santa Catarina, southern Brazil. PeerJ, 4 (e 2629), 1 - 19. https: // doi. org / 10.7717 / peerj. 2629","Monteiro, J. P. C., Condez, T. H., Garcia, P. C. A., Comitti, E. J., Amaral, I. B. & Haddad, C. F. B. (2018 a) A new species of Brachycephalus (Anura, Brachycephalidae) from the coast of Santa Catarina State, southern Atlantic Forest, Brazil. Zootaxa, 4407 (4), 483 - 505. https: // doi. org / 10.11646 / zootaxa. 4407.4.2","Ribeiro, L. F., Blackburn, D. C., Stanley, E. L., Pie, M. R. & Bornschein, M. R. (2017) Two new species of the Brachycephalus pernix group (Anura: Brachycephalidae) from the state of Parana, southern Brazil. PeerJ, 5, e 3603. https: // doi. org / 10.7717 / peerj. 3603","Pie, M. R., Ribeiro, L. F., Confetti, A. E., Nadaline, M. J. & Bornschein, M. R. (2018) A new species of Brachycephalus (Anura: Brachycephalidae) from southern Brazil. PeerJ, 6 (e 5683), 1 - 27. https: // doi. org / 10.7717 / peerj. 5683","Izecksohn, E. (1971) Novo genero e nova especie de Brachycephalidae do Estado do Rio de Janeiro, Brasil (Amphibia, Anura). Boletim do Museu Nacional, Boletim do Museu Nacional (N. S.), Zoologia, New Series, Zoologia, 280, 1 - 12.","Giaretta, A. A. & Sawaya, R. J. (1998) Second species of Psyllophryne (Anura: Brachycephalidae). Copeia, 1998 (4), 985 - 987. https: // doi. org / 10.2307 / 1447345","Napoli, M. F., Caramaschi, U., Cruz, C. A. G. & Dias, I. R. (2011) A new species of flea-toad, genus Brachycephalus Fitzinger (Amphibia: Anura: Brachycephalidae), from the Atlantic rainforest of southern Bahia, Brazil. Zootaxa, 2739 (1), 33 - 40. https: // doi. org / 10.11646 / zootaxa. 2739.1.3","Condez, T. H., Monteiro, J. P. C., Comitti, E. J., Garcia, P. C. A., Amaral, I. B. & Haddad, C. F. B. (2016) A new species of flea-toad (Anura: Brachycephalidae) from southern Atlantic Forest, Brazil. Zootaxa, 4083 (1), 40 - 56. https: // doi. org / 10.11646 / zootaxa. 4083.1.2","Pombal, J. P. Jr. & Izecksohn, E. (2011) Uma nova especie de Brachycephalus (Anura: Brachycephalidae) do estado do Rio de Janeiro. Papeis Avulsos de Zoologia, 51 (28), 443 - 451. https: // doi. org / 10.1590 / S 0031 - 10492011002800001","Guimaraes, C. da S., Luz, S., Rocha, P. C. & Feio, R. N. (2017) The dark side of pumpkin toadlet: a new species of Brachycephalus (Anura: Brachycephalidae) from Serra do Brigadeiro, southeastern Brazil. Zootaxa, 4258 (4), 327 - 344. https: // doi. org / 10.11646 / zootaxa. 4258.4.2","Pombal, J. P. Jr. & Gasparini, J. L. (2006) A new Brachycephalus (Anura: Brachycephalidae) from the Atlantic Rainforest of Espirito Santo, southeastern Brazil. South American Journal of Herpetology, 1 (2), 87 - 93. https: // doi. org / 10.2994 / 1808 - 9798 (2006) 1 [87: ANBABF] 2.0. CO; 2","Condez, T. H., Clemente-Carvalho, R. B. G., Haddad, C. F. B. & dos Reis, S. F. (2014) A new species of Brachycephalus (Anura: Brachycephalidae) from the highlands of the Atlantic Forest, Southeastern Brazil. Herpetologica, 70 (1), 89 - 99. https: // doi. org / 10.1655 / HERPETOLOGICA-D- 13 - 00044","Alves, A. C. R., Sawaya, R., Reis, S. F. & Haddad, C. B. F. (2009) A new species of Brachycephalus (Anura: Brachycephalidae) from the Atlantic Rain Forest in Sao Paulo State, Southeastern Brazil. Journal of Herpetology, 43 (2), 212 - 219. https: // doi. org / 10.1670 / 0022 - 1511 - 43.2.212","Bornschein, M. R., Ribeiro, L. F., Rollo Jr., M. M., Confetti, A. E. & Pie, M. R. (2017) Advertisement call of Brachycephalus albolineatus (Anura: Brachycephalidae). PeerJ, 6 (e 5273), 1 - 18. https: // doi. org / 10.7287 / peerj. preprints. 3461 v 1","Monteiro, J. P. C., Condez, T. H., Garcia, P. C. A. & Haddad, C. F. B. (2018 b) The advertisement calls of two species of Brachycephalus (Anura: Brachycephalidae) from southern forest Brazil. Zootaxa, 4415 (1), 183 - 188. https: // doi. org / 10.11646 / zootaxa. 4415.1.10","Wistuba, E. M. (1998) Historia natural de Brachycephalus pernix Pombal, Wistuba and Bornschein, 1998 (Anura) no Morro Anhangava, Municipio de Quatro Barras, Estado do Parana. M. Sc. dissertation, Universidade Federal do Parana, Curitiba, 89 pp.","Pombal, J. P. Jr., Sazima, I. & Haddad, C. F. B. (1994) Breeding behavior of the pumpkin toadlet, Brachycephalus ephippium (Brachycephalidae). Journal of Herpetology, 28 (4), 516 - 519. https: // doi. org / 10.2307 / 1564972","Araujo, C. B., Guerra, T. J., Amatuzzi, M. C. O. & Campos, L. A. (2012) Advertisement and territorial calls of Brachycephalus pitanga (Anura: Brachycephalidae). Zootaxa, 3302 (1), 66 - 67. https: // doi. org / 10.11646 / zootaxa. 3302.1.5","Tandel, M. C. F. F., Loibel, S., Oliveira, E. G. & Haddad, C. F. B. (2014) Diferenciacao de 3 tipos de vocalizacoes (cantos) na especie Brachycephalus pitanga. Revista da Estatistica da Universidade Federal de Ouro Preto, 3, 374 - 386.","Goutte, S., Mason, M. J., Christensen-Dalsgaard, J., Montealegre-Z, F., Chivers, B. D., Sarria-S, F. A., Antoniazzi, M. M., Jared, C., Sato, L. A. & Toledo, L. F. (2017) Evidence of auditory insensitivity to vocalization frequencies in two frogs. Scientific Reports, 7 (12121), 1 - 9. https: // doi. org / 10.1038 / s 41598 - 017 - 12145 - 5","Verdade, V. K., Rodrigues, M. T., Cassimiro, J., Pavan, D., Liou, N. & Lange, M. C. (2008) Advertisement call, vocal activity, and geographic distribution of Brachycephalus hermogenesi (Giaretta and Sawaya, 1998) (Anura: Brachycephalidae). Journal of Herpetology, 42 (3), 542 - 549. https: // doi. org / 10.1670 / 07 - 287.1","Bokermann, W. C. A. (1966) Lista anotada das localidades tipo de anfibios brasileiros. Servico de Documentacao, Sao Paulo, 183 pp.","Ihering, H. V. & Ihering, R. V. (1911) O Museu Paulista nos anos de 1906 a 1909. Revista do Museu Paulista, 8, 1 - 22. https: // doi. org / 10.5962 / bhl. part. 20998","Campos, L. A., Silva, H. R. & Sebben, A. (2010) Morphology and development of additional bony elements in the genus Brachycephalus (Anura: Brachycephalidae). The Biological Journal of the Linnean Society, 99 (4), 752 - 767. https: // doi. org / 10.1111 / j. 1095 - 8312.2010.01375. x","Alberch, P. & Gale, E. A. (1985) A developmental analysis of an evolutionary trend: digital reduction in amphibians. Evolution, 39, 8 - 23. https: // doi. org / 10.1111 / j. 1558 - 5646.1985. tb 04076. x","Fabrezi, M. (2001) A survey of prepollex and prehallux variation in anuran limbs. Zoological Journal of the Linnean Society, 131 (2), 227 - 248. https: // doi. org / 10.1111 / j. 1096 - 3642.2001. tb 01316. x","Andersen, M. L. (1978) The comparative myology and osteology of the carpus and tarsus of selected anurans. Ph. D. dissertation, University of Kansas, Lawrence, Kansas, 295 pp."]}

Brachycephalus bufonoides was described as a “variety” of B. ephippium based on two specimens which 90 years later was considered full species. Besides its brief original description, nothing else is known for this species. Herein we report the rediscovery of the pumpkin-toadlet Brachycephalus bufonoides from Nova Friburgo, State of Rio de Janeiro, the second most populous area within the Atlantic Forest in Brazil. A detailed osteological description of this species was also provided, including skull, hyolaryngeal skeleton and postcranium skeleton. The laryngeal skeleton of Brachycephalus genus was depicted for the first time. We conducted a molecular phylogenetic analysis of Brachycephalus using DNA sequences comprising two fragments of mitochondrial gene (16S). Both analysis with Bayesian inference and maximum parsimony supported the recognition of B. bufonoides as an exclusive lineage, allocated within the B. ephippium species group in B. vertebralis lineage. We improved the diagnosis and variation of the species, including more collected specimens, coloration in vivo and advertisement call description. Compared with its congeners, B. bufonoides has skin on head and dorsum with dermal hyperossification; skull with hyperossification of postorbital crests; a pair of hyperossified bulges about equidistant between postorbital crests; fourth presacral vertebra with transverse process hyperossified, ornamented and sacral diapophyses hyperossified, which can be seen externally (lineage of B. vertebralis sensu Condez et al. 2020); presence of dermal ossification as separated bulges of each vertebrae; general background color orange with different intensities of dark orange blotches on dorsum, including bordering of sacral region; absence of osteoderms and presence of warts on the dorsolateral surface of body; medium body size (SVL of adults: 12.0–14.5 mm for males and 14.7–16.3 mm for females; Table 1); rough dorsum; advertisement calls with 13 to 17 pulses; presence of pulse period modulation; and advertisement calls with notes longer than 0.2 s (0.22 to 0.31 s). Herein an important contribution for the taxonomy and systematics of this genus is provided, including a large amount of novel information for B. bufonoides from different sources (i.e., molecular, morphological variation, bioacoustic), allowing it to be included in future studies of species delimitation and relationships within Brachycephalus. Also, the discovery of this species reiterates the importance of Nova Friburgo for the conservation of the Atlantic Forest biodiversity.

Made available in DSpace on 2020-12-10T19:51:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundacao Grupo Boticario de Protecao a Natureza Evolutionary changes towards a miniaturized body plan may directly affect other important phenotypic traits related to the physiology, behaviour and ecology of organisms. The frog genus Brachycephalus is an outstanding example of a radiation of miniaturized species endemic to the Brazilian Atlantic Forest. We inferred ancestral states and historical changes in body size, body colour and hyperossification to test hypotheses about diversification and selective environmental mechanisms leading to the evolution of these specialized traits. The ancestral distribution was associated with high-elevation regions in the northern Serra do Mar mountain range, and diversification in the genus was coincident with important geological and climatic events during the history of the Atlantic Forest. The dynamic historical changes provided an opportunity for multiple lowland lineages and for speciation via dispersal and vicariance in multiple invasions of the highlands. The ancestral Brachycephalus was reconstructed as miniaturized and dull coloured, without hyperossification in the skin, skull or postcranial skeleton. A parallel evolution of phenotypic traits has occurred in northern and southern Atlantic Forest lineages, beginning in the Miocene. Shifts in body size are not related to elevation range or latitude. However, we found a significant correlation between the evolution of hyperossification and aposematism with increasing body size. INMA, Ave Jose Ruschi 4, BR-29650000 Santa Teresa, ES, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Zool, Ave 24-A,1515,Caixa Postal 199, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura CAUNESP, Ave 24-A,1515,Caixa Postal 199, BR-13506900 Rio Claro, SP, Brazil Cornell Univ, EEB, Corson Hall, Ithaca, NY 14853 USA Univ Estadual Paulista, Inst Biociencias, Dept Zool, Ave 24-A,1515,Caixa Postal 199, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura CAUNESP, Ave 24-A,1515,Caixa Postal 199, BR-13506900 Rio Claro, SP, Brazil CNPq: 141716/2010-0 CNPq: 201033/2012-7 CNPq: 401729/20133 CNPq: 302518/2013-4 CNPq: 302308/2019-9 FAPESP: 2013/50741-7 FAPESP: 2014/50342-8 FAPESP: 2015/00641-1 FAPESP: 2016/16871-9

When Brachycephalus albolineatus was described, it was known only from its type locality in the state of Santa Catarina, Brazil. Here we provide a new record for the species. Collected specimens were identified in comparison with the type series. We collected 7 specimens in Morro do Garrafão, municipality of Corupá, state of Santa Catarina, at 500–530 m above sea level, in December 2016. The new record is 22 km away from the type locality and also extends the altitudinal distribution of the species, which was previously between 790–835 m above sea level.

Made available in DSpace on 2020-12-12T01:49:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Universidade Estadual Paulista We describe a new species of Ischnocnema from the Serra da Bocaina mountain range, state of São Paulo, southeastern Brazil, based on morphological, bioacoustic, and mtDNA data. The new species is retrieved with high support values within the I. lactea species series as the sister species of I. spanios. Ischnocnema bocaina sp. nov. is characterized by its medium size (18.6–19.0 mm), a smooth venter, a rounded snout in dorsal view and acuminate in lateral view, a slightly expanded subgular, single vocal sac, a round and whitish, poorly-developed glandular-appearing nuptial pad on the dorsal surface of the thumb, and a nonpulsed advertisement call with 9 to 18 notes. We raise to 38 the number of Ischnocnema species, the 12th described in the past 10 years. Instituto de Biociências UNESP – Universidade Estadual Paulista Câmpus Rio Claro Departamento de Zoologia e Centro de Aquicultura (CAUNESP), Cx. Postal 199 Instituto de Ciências Biológicas UFMG – Universidade Federal de Minas Gerais Departamento de Zoologia Laboratório de Herpetologia, Avenida Antônio Carlos, 6627, Pampulha Museu de Zoologia da Universidade de São Paulo, Avenida Nazaré 481 Instituto de Biociências UNESP – Universidade Estadual Paulista Câmpus Rio Claro Departamento de Zoologia e Centro de Aquicultura (CAUNESP), Cx. Postal 199

Advertisement calls can be used to aid solving taxonomic problems and understanding the evolution of certain groups. In this study, the advertisement call of Eleutherodactylus bilineatus is described. It is composed by two different notes with a total duration of 0.529–4.241 seconds and dominant frequency of 1.72–3.45 kHz. Additionally, new data is provided on the geographical distribution of Eleutherodactylus bilineatus and the most inland record for this species.

Made available in DSpace on 2018-12-11T17:22:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-03 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) We report the mitogenomes for five species of the Ischnocnema guentheri series, being the first described for this genus of brachycephalid frogs. We assembled mitogenomes from anchored hybrid enrichment data and recovered the 13 protein-coding genes, 22 tRNA genes, and two rRNA genes for all species. The general structure agrees with most previously sequenced neobatrachians, with two exceptions: the origin of replication of L-strand (OL) was found between tRNA-A and tRNA-N, and the position of tRNA-L and tRNA-T, which are dispersed in the control region. We provide a phylogenetic tree with outgroups, which is consistent with previous phylogenetic hypotheses. Department of Zoology and Aquaculture Center (CAUNESP) Biosciences Institute São Paulo State University–UNESP Department of Zoology Instituto de Biologia Roberto Alcântara Gomes Rio de Janeiro State University–UERJ Department of Scientific Computing Florida State University Department of Biological Science Florida State University Zoological Institute Technical University of Braunschweig Department of Zoology and Aquaculture Center (CAUNESP) Biosciences Institute São Paulo State University–UNESP CNPq: #88881.062205/2014-01 CNPq: 302518/2013-4

Made available in DSpace on 2022-04-28T19:28:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Chico Mendes de Conservação da Biodiversidade Brachycephalus actaeus Monteiro, Condez, Garcia, Comitti, Amaral & Haddad, 2018 is a recently described member of the B. pernix species group from northeastern Santa Catarina, southern Brazil, where it has been recorded from six localities at altitudes of 20–220 m. At the time of its description, this species was believed to be associated with lowlands, a unique trait for members of the B. pernix group. Here, we extend altitudinal distribution of B. actaeus to 530 m elevation and determine that the species occupies only montane forests. Instituto de Biociências Universidade Estadual Paulista, Praça Infante Dom Henrique s/no, Mater Natura – Instituto de Estudos Ambientais, Rua Lamenha Lins 1080 Escola de Ciências da Vida Pontifícia Universidade Católica do Paraná, Rua Imaculada Conceição 1155, Curitiba Departamento de Zoologia Universidade Federal do Paraná, Caixa Postal 19020 Instituto de Biociências Universidade Estadual Paulista, Praça Infante Dom Henrique s/no, FAPESP: 17/21611-9 CAPES: 301636/2016-8571334/2008–3 CNPq: 301636/2016-8571334/2008–3

Made available in DSpace on 2020-12-10T19:35:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-09-13 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Minas Gerais Research Foundation Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Grupo O Boticario de Protecao a Natureza Instituto Mater Natura de Estudos Ambientais Instituto Neotropical de Pesquisa e Conservacao Bioacoustics is a powerful tool used for anuran species diagnoses, given that advertisement calls are signals related to specific recognition and mate attraction. Thus, call descriptions can support species taxonomy. In spite of that, call descriptions are lacking for many species, delaying advances in biodiversity research. Here, we describe the advertisement calls of 20 anuran species from the Brazilian Atlantic Forest. We accessed 50 digital recordings deposited in the Fonoteca Neotropical Jacques Vielliard. Acoustic analyses were carried out in the software Raven pro 1.5. We provide a general comparison of call structure among species inside taxonomic groups and genera. The vocalizations described here belong to poorly known species, which are representatives of six families: Brachycephalidae, Bufonidae, Ceratophryidae, Cycloramphidae, Hylidae, and Phyllomedusidae. Despite this, still there are 163 species of anurans from Atlantic Forest with calls not formally described. Our work represents an important step in providing data for a taxonomic perspective and improving the knowledge of the Atlantic Forest anuran diversity. Univ Fed Bahia, Inst Biol, Salvador, BA, Brazil Univ Estadual Paulista, Dept Zool, Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura CAUNESP, Rio Claro, SP, Brazil Univ Fed Vicosa, Inst Ciencias Biol & Saude, Florestal, MG, Brazil Univ Estadual Norte Fluminense, Ctr Biociencias & Biotecnol, Campos Dos Goytacazes, RJ, Brazil Univ Fed Vicosa, Programa Posgrad Biol Anim, Vicosa, MG, Brazil Univ Fed Parana, Dept Zool, Curitiba, Parana, Brazil Univ Fed Parana, Programa Posgrad Zool, Curitiba, Parana, Brazil Univ Fed Minas Gerais, Dept Zool, Belo Horizonte, MG, Brazil Univ Estadual Campinas, Dept Biol Anim, Campinas, SP, Brazil Univ Estadual Paulista, Dept Zool, Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura CAUNESP, Rio Claro, SP, Brazil FAPESP: 2013/21519-4 FAPESP: 2016/25358-3 FAPESP: 2013/50741-7 FAPESP: 2014/50342-8 CNPq: 300896/2016-6 CNPq: 438675/2016-9 CNPq: 307091/2015-5 CNPq: 302518/2013-4 Minas Gerais Research Foundation: FAPEMIG-APQ-01796-15 FAPEMIG: RDP-00004-17 Grupo O Boticario de Protecao a Natureza: 085420101/A0001_2012

Made available in DSpace on 2020-12-12T00:54:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Brachycephalus fuscolineatus Pie, Bornschein, Firkowski, Belmonte-Lopes & Ribeiro, 2015 was described from Morro do Baú, Santa Catarina, Brazil. This species occurs at elevations of 640–790 m, and it is known to have an extent of occurrence of 23.63 ha. Here, we report a new record of B. fuscolineatus from Morro Braço da Onça, Luiz Alves, Santa Catarina. We recorded it on 21–22 December 2018 at elevations between 525 and 530 m; the new record occupied an area of 0.19 ha. We estimate 9 calling males in 36.35 m2. Instituto de Biociências Universidade Estadual Paulista (UNESP), Praça Infante Dom Henrique s/no Instituto de Estudos Ambientais, Rua Lamenha Lins 1080 Escola de Ciências da Vida Pontifícia Universidade Católica do Paraná Curitiba Instituto de Biociências Universidade Estadual Paulista (UNESP), Praça Infante Dom Henrique s/no

Most miniaturized froglets of the genus Brachycephalus occur in isolation in slopes of mountain ranges at elevations varying from 600 to 1,800 m in the Atlantic Forest of eastern Brazil. For organisms such as Bracycephalus with spatially discontinuous distributions, a fundamental task is to determine whether observed patterns of variation are consistent with geographic differentiation among allopatric populations within a single species or are suggestive of a potential species boundary. We address this problem by focusing on continental and island population samples potentially assignable to Brachycephalus nodoterga (Anura: Brachycephalidae) from the perspective of variation in qualitative and quantitative morphological traits and DNA sequences. Population samples from continental and island populations share color characteristics, qualitative traits, and multivariate patterns of variation and covariation in cranial metric traits. Comparative analysis of DNA sequences showed the magnitude of molecular distances between B. nodoterga and Brachycephalus ephippium to be 1 order of magnitude larger than molecular distances within B. nodoterga and B. ephippium. We interpret the combined morphological and molecular evidence to indicate that continental and island population samples examined here are conspecific. Therefore, by defining species boundaries for B. nodoterga, we also established minimal estimates of its distribution. Fil: Clemente Carvalho, Rute B. G.. Universidade Vila Velha; Brasil Fil: Perez, Sergio Ivan. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina Fil: Tonhatti, Carlos H.. Universidade Estadual de Campinas; Brasil Fil: Condez, Thais H.. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil Fil: Sawaya, Ricardo J.. Universidade de Sao Paulo; Brasil Fil: Haddad, Celio F. B.. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil Fil: Dos Reis, Sergio F.. Universidade Estadual de Campinas; Brasil

Made available in DSpace on 2019-10-04T12:31:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-03 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundacao Grupo Boticario de Protecao a Natureza A new miniaturized frog of the genus Brachycephalus (Anura: Brachycephalidae) i s described from Morro Santo Anjo in the municipality of Massaranduba, Santa Catarina, southern Brazil. Specimens were collected from the leaf litter between 470 and 540 above sea level. The new species is distinguished from all its congeners by the combination of the following characters: (1) body robust and bufoniform; (2) size snout-vent length 9.9-11.7 mm for males and 10.0-12.9 mm for females; (3) smooth dorsum; (4) general color (in life) orange with white dots and stripe in the middle of the head and along its vertebral column; (5) iris completely black; (6) advertisement call composed of note groups; (7) isolated notes with 1-3 pulses; and (8) short isolated notes (0.002-0.027 s). An estimate of the male density of the new species is also presented. Phylogenetic information indicates that the new species is part of the southernmost Glade of Brachycephalus, which includes Brachycephalus fuscolineatus, B. albolineatus, and B. boticario. The severe anthropogenic impacts in and around the type locality indicate that immediate actions should be taken to ensure the long-term preservation of the new species. Univ Fed Parana, Dept Zool, Curitiba, Parana, Brazil Mater Nat Inst Estudos Ambientais, Curitiba, Parana, Brazil Pontificia Univ Catolica Parana, Escola Ciencias Vida, Curitiba, Parana, Brazil Univ Fed Parana, Programa Posgrad Zool, Curitiba, Parana, Brazil Univ Estadual Paulista, Inst Biociencias, Campus Litoral Paulista, Sao Vicente, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Campus Litoral Paulista, Sao Vicente, SP, Brazil CNPq: 301636/2016-8 Fundacao Grupo Boticario de Protecao a Natureza: A0010_2014

Ischnocnema lactea (Miranda-Ribeiro 1923) Basanitia lactea Miranda-Ribeiro 1923: 851 Eleutherodactylus lacteus —Lynch 1968: 875 Eleutherodactylus (Eleutherodactylus) lacteus — Lynch & Duellman 1997: 226 Ischnocnema lactea —Heinicke et al. 2007: 15; Hedges et al. 2008: 27 Lectotype. MZUSP 828; adult female, collected by Gustavo Edwall from the Municipality of Iguape, State of São Paulo, Brazil. The original description did not provide a date or specific sampling locality (see discussion for an analysis on the type locality). Bokermann (1966) designated the lectotype. Paralectotype. MZUSP 504; adult male of Ischnocnema parva (Girard 1853) from Campo Grande, Municipality of Santo André, State of São Paulo, Brazil. The paralectoype was designated by Bokermann (1966) and later redetermined by Heyer (1985) as Ischnocnema parva. Type locality. Municipality of Iguape, State of São Paulo, Brazil. Restricted by Bokermann (1966). We note, however, that the record for the Municipality of Iguape, which was formerly considered the type locality, is in error. Here, we redefine the type locality as the area surrounding the Padre Dória road, in the Juqueriquerê River basin (coordinates 23°38'32.43"S, 45°41'39.20"W, datum = WGS84; altitude 1,010 m a.s.l.), Municipality of Caraguatatuba, State of São Paulo, Brazil (see discussion for details). Examined specimens. A total of 47 specimens of I. lactea were examined, which included 18 adult males, 13 adult females (including the lectotype; Fig. 2), 13 sub-adults, two young individuals, and one adult of undetermined sex. Specimens were obtained from different populations inhabiting the Serra do Mar region in the State of São Paulo, Southeastern Brazil: Núcleo Curucutu, PESM, Municipality of Itanhaém (N= 7), and Municipality of São Paulo (N= 1); Núcleo Santa Virgínia, PESM, Municipality of São Luís do Paraitinga (N= 1); Serra da Cantareira, Municipality of São Paulo (N= 1); Reserva Florestal do Morro Grande, Municipality of Cotia (N= 2); Estação Biológica de Boracéia (EBB), Municipality of Salesópolis (N= 2); PNMNP, Municipality of Santo André (N= 21); Parque Estadual de Ilhabela (PEI), Municipality of Ilhabela (N= 2), and Municipality of Iguape (type locality; N= 1, lectotype). Juveniles (MZUSP 134037, 145253) and sub-adults (four males, CFBH 11171, 16772, 29118, ZUEC 6382; nine females, CFBH 17670, 29094, 29107-08, MZUSP 145260, ZUEC 3531, 6381, 6383, 6470) were not considered in morphological and morphometric comparisons. Moreover, lectotype MZUSP 878 was in moderate condition of preservation (Fig. 2). Diagnosis. Ischnocnema lactea is diagnosed by the following combination of characters: (1) large-sized species (among I. lactea species), with adult males SVL 19.6–26.7 mm; X = 23.3. ± 2.4 mm; N = 18; adult females SVL 27.6–34.3 mm; X = 30.2 ± 2.5 mm; N = 10; (2) dorsum, lateral, and gular region rugose with several scattered tubercles; (3) venter moderately granular; (4) head longer than wide; (5) snout rounded in dorsal view; (6) tip of snout obtuse in lateral view; (7) eyelid tubercles oval, well-developed; (8) vomerine process present, bearing prevomerine odontoids; (9) vocal sac single, subgular, externally expanded; (10) ungual flap present, disks expanded and indented, T-shaped, truncate in fingers II–IV; (11) double glandular nuptial pad; (12) toes relative length IIschnocnema lactea. Ischnocnema lactea is distinguishable from all other species of I. lactea as follows (character states for other members in parentheses): large-sized species with adult males reaching SVL 19.6–26.7 mm, and adult females SVL 27.6–34.3 mm (adult males are smaller in I. concolor 14.7–18.4 mm, I. gehrti 14.2–15.3 mm, I. melanopygia 15.7–18.7 mm, I. paranaensis 17.7 mm, I. randorum 11.8– 15.0 mm, I. spanios 14.7 mm, and I. vizottoi 13.3–17.0 mm; overlap in size on adult males of I. holti 18.4–24.9 mm and I. nigriventris 18.2–20.2 mm); dorsum very rugose, with several tubercles and granules (dorsum smooth in I. concolor, I. gehrti, I. melanopygia, I. paranaensis, I. randorum, and I. spanios); snout rounded in dorsal profile (subovoid in I. gehrti, I. melanopygia, I. randorum, I. spanios, and I. vizottoi; rounded in the other species); tip of snout may present a medial process, providing a subovoid aspect in dorsal view (variation also occurs in I. concolor and I. vizottoi); tip of snout obtuse in lateral view (also obtuse in I. holti; truncate in I. gehrti and I. spanios; rounded in other species); eyelid tubercles oval well developed (absent in I. concolor; oval poorly developed in I. gehrti, I. holti, I. melanopygia, I. randorum, and I. spanios; conical well developed in I. nigriventris; unknown in I. paranaensis); patches of prevomerine odontoids present (absent in I. paranaensis and I. randorum); vocal sac single, subgular and externally expanded (not expanded in I. gehrti, I. holti, I. spanios, and I. melanopygia; unknown in I. paranaensis); in life, iris varies from golden to lemon-yellowish, with a medial stripe varying from reddish to orange, crossing the black pupil (golden to lemon-yellowish, with brown to black medial stripe in I. concolor, I. holti, I. melanopygia, I. randorum, and I. vizottoi; iris silver with a ventral yellowish copper band in I. nigriventris; dorsally bluish in I. spanios; not reported for I. gehrti and I. paranaensis); disks expanded, T-shaped, truncate in fingers II–IV (similar for most species; disks expanded Tshaped, truncate in fingers III–IV in I. concolor, I. melanopygia, and I. vizottoi; disk finger II rounded in I. melanopygia, I. vizottoi and, I. concolor, slightly expanded in the latter); double glandular nuptial pads present (single glandular in I. concolor, I. gehrti, I. melanopygia, I. randorum, I. spanios, and I. vizottoi; unknown in I. paranaensis); toes relative length II. gehrti, I. melanopygia and I. spanios); calcar tubercles moderate (also moderate in I. randorum, I. spanios, and I. gehrti; poorly developed in I. holti, I. concolor, and I. vizottoi, occasionally inconspicuous in the last two species; well developed in I. melanopygia and I. nigriventris; absent in I. paranaensis); in life, inguinal region and hidden areas of thigh, shank, and tarsus show a large array of bright colors and patterns (bright coloration on mentioned areas absent in I. concolor, I. paranaensis, and I. vizottoi); bright colors on mentioned areas appear on marbled pattern in a dark background (pattern does not occur in I. concolor, I. gehrti, I. paranaensis, I. randorum, and I. spanios); black blotch on cloacal region, tarsus, and feet present (absent in most of the species, except in I. gehrti, I. melanopygia, and I. spanios; unknown for I. paranaensis). Redescription of the lectotype MZUSP 828. According to original description (Miranda-Ribeiro 1923), adult female from the Municipality of Iguape, State of São Paulo, Southeastern Brazil, collected by Gustavo Edwall. The specimen shows a moderate condition of preservation, having lost some general characters. The lectotype has a robust body, dorsum smooth, presenting very few small, scattered tubercles; flanks and ventral surfaces granular (bely and laterals) to very granular (cloacal region); skin translucent, with muscles seen by transparency mainly on arms and legs. Venter almost entirely open with organs exposed; pectoral and abdominal folds indistinct. Head slightly longer than wide, head length 37% of SVL; snout acuminate in lateral view and subovoid in dorsal; upper jaw show a large medial cut, and projects beyond lower jaw; large eyes, anterolaterally oriented; pupil horizontal and elliptical; translucent palpebral membrane, faintly pigmented on edge; both eyelids present some tuberculation mainly on edges; tympanum moderately distinct, with membrane hidden beneath skin and annulus evident only ventrally; supratympanic fold poorly developed, probably shabby; three medium to large-sized postcommissural tubercles on both side of head. Canthus rostralis distinct, straight; loreal region oblique, dorsolaterally directed and slightly concave; nostrils oval, protruding in dorsal and lateral views, posterolaterally directed; inter-nostril distance 22.7% of HW; choanae nearly rounded, distant from one; prevomerine processes with odontoids present, oblique, situated between and posterior to the choanae; thick, medium-sized tongue, elliptically disposed on mouth floor, notched medially. Arms and forearms moderately slender. Fingers long, with all finger tips lost, which does not allow comparisons with the original state; subarticular tubercles single, oval, well developed; outer metacarpal tubercle cordiform and inner metacarpal tubercle oval, both worn out; supernumerary tubercles on palmar region absent or inconspicuous. Legs moderately slender; thigh almost as long as shank; feet with worn conical outer metatarsal tubercle, and elliptical inner metatarsal tubercle with detached external edge; both inner and external metatarsal tubercles moderately developed; subarticular oval tubercles moderately developed; supernumerary tubercles absent or inconspicuous. Toes long, with some toes tips lost, not allowing to state their original relative length; toes not webbed nor fringed, with ungual flaps present; toes I–V with disk expanded, rounded, dorsally emarginated, indented; disk pads even, broadened, following their disks sizes; calcar inconspicuous. Measurements of lectotype MZUSP 828 (in mm). SVL 30.5; HL 11.4; HW 11.3; ED 4.4; END 2.8; ESD 4.9; IOD 2.8; IND 2.6; TD 1.3; FAL 6.5; HAL 7.5; THL 13.8; SHL 15.3; FL 19.0, and 4TL 7.8, both considering the lost of 4TD. Color in preservative. General color of body varying from cream to milky. Few blotches, dots, and stripes can still be observed along the body surfaces. Some light brown blotches are present on dorsum, laterals, head, and dorsal region of thighs. Intraspecific variation based on newly collected adult specimens. In general, specimens show a body robust, dorsum very rugose, presenting several tubercles of varied sizes. Flanks and undersurfaces varied from granular (venter, arms, and legs) to very granular (lateral, cloacal, and gular regions). In comparison to live individuals, fixed specimens might present body surfaces much less rugose and granular, probably due to fixation. Rugosity also varies in intensity on different parts of the body of the same specimen. Pectoral fold tenuous, lined from axilla to axilla, positioned slightly below the arm–axilla joint; abdominal fold distinct, more conspicuous at the posterior portion of belly than laterals. Head longer than wide, snout acuminate in lateral view, and subovoid in dorsal view mainly due to a medium to large tubercle that can be present at the tip of snout (e.g., CFBH 29055; MZUSP 145317); upper jaw projecting beyond lower jaw; large eyes, protuberant when opened, anterolaterally oriented, with pupil horizontal and elliptical; translucent palpebral membrane, pigmented on edge; eyelids vary from presenting none to several oval tubercles well developed; tympanum moderately distinct, with membrane hidden beneath skin and annulus evident only ventrally; supratympanic fold moderately developed; three to four medium to large-sized postcommissural conical tubercles can be present on both side of head. There are some small to medium-sized conical tubercles that form the end of supratympanic fold. Canthus rostralis distinct, straight; loreal region oblique, dorsolaterally directed and slightly concave; nostrils oval, protruding in dorsal and lateral views, posterolaterally directed; inter-nostril distance average 27.1% of HW; choanae nearly rounded, distant from one another; prevomerine processes present, oblique, well developed (generally twice its width), situated between and posterior to the choanae, with about four to six odontoids on each patch; premaxillary teeth present; thick, medium-sized tongue, elliptically disposed, notched medially; vocal slits large, conspicuous, anterolaterally oriented, beginning anteriorly to the tongue notch, and ending anteriorly to the line of buccal commissure. Vocal sac single, largely expanded, covering all over the gular region, dorsally visible, however, some males may show vocal sacs not expanded (e.g., CFBH 1936, 28167) and, in general, the gular region of males is darker than in females. Some adult males with a dark gular region may also show a lighter medial line (e.g., CFBH 19368, 29055, 29082). Arms and forearms moderately robust; three to five lined-up tubercles moderately developed are present on ventral surface of each forearm. Fingers long, not fringed, with ungual flaps present; relative length of fingers IColor in life (Fig. 5). Despite of the general milky color of lectotype, which led Alípio de Miranda-Ribeiro to choose the epithet ‘lactea’ (lacteus = milk), our results show that specimens of Ischnocnema lactea have stunningly attractive colorful patterns that display a large array of bright colors, mainly on hidden areas, with a high level variation on life coloration. The dorsum presents several blotches and/or large stripes that vary in position, dimension, shapes, colors, and intensity of tones, contrasting with background general color. An interocular blotch is usually present, being medially divided in some specimens; two small blotches between eyes and nostrils can be present, being connected as a stripe in some individuals; black supratympanic stripe conspicuous. The dorsum background color varies in brown, ranging from dark to light brown, nut-brown, orange, green, and grayish; some individuals also showed two or more colors on dorsal background. On flanks, colors fade into lighter variants of those colors, which may become cream nearby ventral surface. Forearms may exhibit two to three bars that can be faint or poorly conspicuous in some specimens; a blotch of varying color may be present on elbow. Thighs, shanks, and tarsus plus foot have three to four bars (each) that can be faint or inconspicuous in some specimens. Venter and hidden areas of inguinal region, thighs, shanks, and tarsus also present a large array of bright colors and patterns; background color ranged from intense to lighter tones of red, yellow, orange, black, gray, and cream/beige. Colors may be uniformly distributed all over the venter or varied in tones and intensity on different regions of undersurfaces, being, for example, lighter on gular and/or chest regions and more intense on belly and legs. The gular region on males can be darker than the rest of venter. The iris is golden with a medial red stripe crossing the black pupil, presenting several black vermiculations mainly in the ventral band. Two females, MZUSP 145318 and CFBH 25822, presented a colored dorsal band orange and green, respectively, externally lined by a dark line, delimiting dorsum from lateral surface. Color in preservative. Background color in preservative faints into different tones of gray and silver. Our observations of old-dated fixed specimens revealed that hidden surfaces become slightly opaque, maintaining the same colors, while patterns and bright colors disappear with time. In general, all specimens maintained the former pattern of blotches, dots, and stripes in all surfaces. Geographic distribution. Ischnocnema lactea is endemic to the Atlantic Rainforest biome, occurring in central portion of Serra do Mar, in the State of São Paulo, Southeastern Brazil. It inhabits mountainous forests at moderate altitudes, occurring from 700–1,120 m a.s.l. (Fig. 6). Coordinates of localities are provided in Appendix 2. Vocalization and acoustic comparisons. The advertisement call of Ischnocnema lactea consists on a single pulsed note (notes emitted in series in I. nigriventris and I. randorum), containing from eight to thirteen pulses (Fig. 7). The mean dominant frequency is 2418.9 ± 17.9 Hz (2239.5–2756.2 Hz, N = 24 notes). The mean note duration is 908 + 144 ms (634–1060, n = 24 notes). The mean of pulses per note is 10.45 + 1.10 (8–13, n = 24 notes), and note rate of 11.01 + 3.77 notes per minute (6.77–14.29, n = 24 notes). The, Published as part of Silva-Soares, Thiago, Malagoli, Leo Ramos, Cruz, Carlos Alberto Gonçalves & Caramaschi, Ulisses, 2018, On the taxonomy and natural history of the secretive Ischnocnema lactea (Miranda-Ribeiro, 1923) (Anura: Brachycephaloidea: Brachycephalidae), pp. 137-153 in Zootaxa 4444 (2) on pages 140-147, DOI: 10.11646/zootaxa.4444.2.3, http://zenodo.org/record/1309568, {"references":["Miranda-Ribeiro, A. (1923) Basanitia lactea (Um novo batracchio das colleccoes do Museu Paulista). Revista do Museu Paulista, 13, 851 - 852.","Lynch, J. D. & Duellman, W. E. (1997) Frogs of the genus Eleutherodactylus in western Ecuador. Systematics, ecology, and biogeography. Miscellaneous Publication. Museum of Natural History, University of Kansas, 23, 1 - 236.","Hedges, S. B., Duellman, W. E. & Heinicke, M. P. (2008) New World direct-developing frogs (Anura: Terrarana): Molecular phylogeny, classification, biogeography, and conservation. Zootaxa, 1737, 1 - 182.","Bokermann, W. C. A. (1966) Lista anotada das localidades tipo de anfibios brasileiros. Sao Paulo, Servico de Documentacao da Universidade de Sao Paulo, Sao Paulo, 183 pp.","Heyer, W. R. (1985) New species of frogs from Boraceia, Sao Paulo, Brazil. Proceedings of the Biological Society of Washington, 98, 675 - 671.","Canedo, C. & Haddad, C. F. B. (2012) Phylogenetic relationships within anuran clade Terrarana, with emphasis on the placement of Brazilian Atlantic rainforest frogs genus Ischnocnema (Anura: Brachycephalidae). Molecular Phylogenetics and Evolution, 65, 610 - 620. https: // doi. org / 10.1016 / j. ympev. 2012.07.016","Frank, N., & Ramus, E. (1995) Complete Guide to Scientific and Common Names of Amphibians and Reptiles of the World. N. G. Publishing Inc., Pottsville, 377 pp."]}

The original description of Ischnocnema lactea (Miranda-Ribeiro, 1923) was based on two specimens and an inaccurate type locality. These circumstances have obscured the identity of this name. Moreover, the I. lactea species has had a complex taxonomic history and, in general, specimens of species in this group are poorly represented in amphibian collections. The diagnosis of the series is confusing, with many non-exclusive and variable characters barely useful. An accurate diagnosis for the series requires a precise characterization of I. lactea, its type species. Recently discovered populations, museum data, and new information on the collector of the type specimen allowed the elucidation of important issues related to this species. Here we provide the redescription of Ischnocnema lactea based on the lectotype, and intraspecific variation based on newly collected specimens. Ischnocnema lactea is characterized by its large body size (adult 19.6–26.7 mm SVL; females 27.6–34.3 mm SVL); dorsum and flanks rugose, venter granular; vocal sac single and subgular, externally expanded; finger disks expanded, T-shaped, indented and truncate in fingers II–IV; toes relative length I

Made available in DSpace on 2018-11-26T17:54:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-20 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundacao Grupo Boticario de Protecao a Natureza Background: Brachycephalus are among the smallest terrestrial vertebrates in the world. The genus encompasses 34 species endemic to the Brazilian Atlantic Rainforest, occurring mostly in montane forests, with many species showing microendemic distributions to single mountaintops. It includes diurnal species living in the leaf litter and calling during the day, mainly during the warmer months of the year. The natural history of the vast majority of the species is unknown, such as their advertisement call, which has been described only for seven species of the genus. In the present study, we describe the advertisement call of Brachycephalus albolineatus, a recently described microendemic species from Santa Catarina, southern Brazil. Methods: We analyzed 34 advertisement calls from 20 individuals of B. albolineatus, recorded between 5 and 6 February 2016 in the type locality of the species, Morro Boa Vista, on the border between the municipalities of Jaragua do Sul and Massaranduba, Santa Catarina, southern Brazil. We collected five individuals as vouchers (they are from the type series of the species). We used the note-centered approach to describe the advertisement calls of the species. Results: B. albolineatus have a long advertisement call of 40-191 s (mean of 88 s) composed of 8-29 notes (mean of 17 notes) emitted at a rate of 6-18 notes per minute (mean of 11 notes per minute) and at a note dominant frequency of five to seven kHz (mean of six kHz). Advertisement calls are composed of isolated notes and note groups (two notes involved in each particular note group); the former is composed by one to three pulses (mean of 2.0) and the note groups by two or three pulses in each note (mean of 2.7). Most advertisement calls present both isolated notes and note groups, with a few cases showing only the former. Note groups are emitted invariably in the last third of the advertisement call. Most isolated notes escalate their number of pulses along the advertisement call (1-2, 1-3 or 2-3). Note duration of isolated notes varies from 0.002 to 0.037 s (mean of 0.020 s) and duration of note group vary from 0.360 to 0.578 s (mean of 0.465 s). Discussion: Individuals increase the complexity of their calls as they proceed, incorporating note groups and pulses per note. Intra-individual variation analysis also demonstrated that less structured advertisement calls (i.e., with notes with fewer pulses) are not stereotyped. It is possible that isolated notes and note groups could have distinct functions, perhaps territorial defense and mating, respectively. We argue that using a note-centered approach facilitates comparisons with calls of congeners, as well as underscores the considerable differences in call structure between species in a single group and among species groups. Univ Estadual Paulista, Inst Biociencias, Sao Vicente, SP, Brazil Mater Nat Inst Estudos Ambientais, Curitiba, Parana, Brazil Pontificia Univ Catolica Parana, Escola Ciencias Vida, Curitiba, Parana, Brazil Univ Fed Parana, Programa Posgrad Zool, Curitiba, Parana, Brazil Univ Fed Parana, Dept Zool, Curitiba, Parana, Brazil Univ Estadual Paulista, Inst Biociencias, Sao Vicente, SP, Brazil CNPq: 301636/2016-8 Fundacao Grupo Boticario de Protecao a Natureza: A0010_2014

The advertisement calls are known for only eight species within the genus Brachycephalus. Herein, we describe the advertisement calls of B. olivaceus and B. quiririensis, both species from southern Atlantic Forest, state of Santa Catarina, Brazil. The advertisement calls of these species are similar, composed by one short and high-frequency note. The parameters range overlap between calls, as exemplified by the dominant frequency of 6.4–7.0 kHz in B. olivaceus and 6.2–6.5 kHz in B. quiririensis. Additionally, we provide a compilation of the available bioacoustic information for this genus.

Brachycephalus actaeus sp. nov. Figures 1, 2, 6, 7, and 8 Holotype. CFBH 39850, adult male, collected at Serra da Palha, Laranjeiras, Ilha de S��o Francisco do Sul, municipality of S��o Francisco do Sul, state of Santa Catarina, Brazil (26��17'50"S; 48 �� 40'28"W, Datum WGS 84, ca 60 meters above sea level), on 19 May 2015, by C.F.B. Haddad, J.P.C. Monteiro, and E.C. Nardin (Figures 1 and 2). Paratopotypes. CFBH 39851, adult female, collected with the holotype; CFBH 39872, 39873, and UFMG 18973, adult males, collected on 25 November 2015, by J.P.C. Monteiro, T.H. Condez, and E.C. Nardin; CFBH 39876, adult male, and CFBH 39877, adult female, collected on 0 2 December 2015, by J.P.C. Monteiro and E.C. Nardin. Paratypes. CFBH 39846, adult male, cleared and double-stained, collected at Fazenda Morro Grande, Morro Grande, Ilha de S��o Francisco do Sul, municipality of S��o Francisco do Sul, state of Santa Catarina, Brazil (26 �� 17'47"S; 48��37'10"W, Datum WGS 84, ca 60 meters above sea level), on 14 November 2014, by E.J. Comitti. UFMG 18970, adult female, collected on 15 November 2014, by J.P.C. Monteiro, T.H. Condez, and E.J. Comitti; CFBH 39849, sub-adult female, collected on 21 November 2014, by J.P.C. Monteiro and E.C. Nardin; CFBH 39848, adult male, cleared and double-stained, collected on 21 January 2015, by C.F.B. Haddad, J.P.C. Monteiro, T.H. Condez, and E.J. Comitti; UFMG 18971, adult male, collected on 0 1 August 2015, by J.P.C. Monteiro and E.C. Nardin; CFBH 39855���39858 and 39861, adult males, CFBH 39860, adult female, CFBH 39859, juvenile, collected on 23 November 2015, by J.P.C. Monteiro, T.H. Condez, and E.C. Nardin; all collected at Centro de Estudos e Pesquisas Ambientais da Univille (CEPA), Vila da Gl��ria, Distrito do Sa��, municipality of S��o Francisco do Sul, state of Santa Catarina, Brazil (26 �� 13'39"S; 48 �� 41'31"W, Datum WGS 84, ca 120 meters above sea level). CFBH 39853 and 39854, adult males, collected on 27 August 2015, by J.P.C. Monteiro and E.C. Nardin; CFBH 39862, adult female, collected on 23 November 2015, by J.P.C. Monteiro, T.H. Condez, and E.C. Nardin; at Estrada do Sa��, Distrito do Sa��, municipality of S��o Francisco do Sul, state of Santa Catarina, Brazil (26��12'06"S; 48 �� 41'37"W, Datum WGS 84, ca 80 meters above sea level). CFBH 39863, 39864, 39867, 39868, and UFMG 18972 adult females, CFBH 39865 and 39870, adult males, collected on 24 November 2015, by J.P.C. Monteiro, T.H. Condez, and E.C. Nardin; CFBH 39875, adult female, collected on 30 November 2015, by J.P.C. Monteiro and E.C. Nardin; at Bra��o do Norte, municipality of Itapo��, state of Santa Catarina, Brazil (26��07'29"S; 48 �� 43'48"W, Datum WGS 84, ca 220 meters above sea level). CFBH 42005���42008, adult females, collected on 17 September 2016, by J.P.C. Monteiro and E.C. Nardin; Fazenda Palmito Juriti, municipality of S��o Francisco do Sul, state of Santa Catarina, Brazil (26��08'09"S; 48 �� 43'54"W, Datum WGS 84, 125���170 meters above sea level). Diagnosis. Brachycephalus actaeus sp. nov. is a new species of the B. pernix group, distinguished from all its congeners by the following combination of characters: (1) ���bufoniform��� body; (2) general dorsal body color dark green with a dark brown vertebral stripe, and orange background more evident in ventral view; (3) absence of hyperossification of the skull and skeleton; (4) pectoral girdle arciferal and robust, with small ovoid fenestra, distant from the epicoracoid; (5) radius and ulna fused; (6) finger IV greatly reduced, almost not visible externally; (7) manus with two prepollical elements; (8) tips of terminal phalangeal elements of fingers I and IV pointed, II and III arrow-shaped; (9) tibiale and fibulare completely fused; (10) toes I and V present but externally indistinguishable, toe II greatly reduced, toe III short and distinct, and toe IV larger and robust; (11) pes with distal tarsal element I present; (12) pes with phalangeal formula 1���2���3���4���0; (13) tips of terminal phalangeal elements of toes I and II pointed, and of toes III and IV arrow-shaped; (14) arytenoid cartilages not mineralized; (15) body size (SVL of adults: 9.2���10.8 mm for males and 11.1���12.4 mm for females); (16) proportional measurements HL/SVL 18���24% and ED/HL 52���73%; (17) rounded snout in dorsal and lateral views; (18) protuberant nostrils; (19) skin texture rough; and (20) advertisement call short (0.02���0.03 seconds), composed of one high-frequency note (dominant frequency 6.6���7.3 kHz). Comparisons with other species. Brachycephalus actaeus sp. nov. exhibits a ���bufoniform��� body and an orange background color, both characteristics that clearly differentiate it from the ���leptodactyliform��� species: B. didactylus, B. hermogenesi, B. pulex, and B. sulfuratus (Izecksohn 1971; Giaretta & Sawaya 1998; Napoli et al. 2011; Condez et al. 2016). These species are generally smaller, exhibit ���leptodactyliform��� bodies, and always exhibit a brown background color (Napoli et al. 2011; Condez et al. 2016). The absence of hyperossification of the skull and skeleton distinguishes Brachycephalus actaeus sp. nov. from B. darkside, B. ephippium, B. garbeanus, and B. margaritatus, which exhibit the extreme condition of hyperossification within Brachycephalus, including a dorsal bony shield (Clemente-Carvalho et al. 2009; Guimar��es et al. 2017). The absence of hyperossification also distinguishes the new species from B. alipioi, B. atelopoide, B. bufonoides, B. crispus, B. guarani, B. nodoterga, B. pitanga, B. toby, and B. vertebralis, which exhibit the intermediate condition of hyperossification within Brachycephalus (Clemente-Carvalho et al. 2009; Haddad et al. 2010; Pombal 2010; Clemente-Carvalho et al. 2011; Condez et al. 2014; Condez et al. 2016). Brachycephalus actaeus sp. nov. differs from B. brunneus, B. coloratus, B. ferruginus, B. izecksohni, and B. pombali, which possess pectoral girdles with larger fenestra, disposed closer to the epicoracoid (small fenestra distant from the epicoracoid in the new species). Likewise, B. brunneus, B. ferruginus, B. izecksohni, and B. pombali differ in having the radius and ulna not fused, pes with distal tarsal element I absent, and toe V reduced (Ribeiro et al. 2005; Alves et al. 2006); the new species has the radius and ulna fused, pes with distal tarsal element I present, and toe V externally not visible. Also, B. albolineatus , B. coloratus, B. curupira, B. ferruginus, and B. pombali have just one prepollical element, and B. izecksohni has no prepollical element (Ribeiro et al. 2005; Alves et al. 2006; Bornschein et al. 2016b; Ribeiro et al. 2017); the new species has two prepollical elements. The phalangeal formula for the pes of B. brunneus , B. izecksohni, and B. pombali is 0���2���3���4���0 (Ribeiro et al. 2005; Alves et al. 2006), and for B. curupira it is 0���1���3���4���0 (Ribeiro et al. 2017); the phalangeal formula of the pes of the new species is 1���2���3���4���0. In B. albolineatus, B. coloratus, and B. curupira the fibulare and tibiale are not completely fused; tips of terminal phalangeal elements of toes II���IV are arrow-shaped (fibulare and tibiale completely fused; tips of terminal phalangeal elements of toes I and II pointed, III and IV arrow-shaped in the new species). Finally, the arytenoid cartilages are mineralized in B. albolineatus and B. coloratus (arytenoid cartilages not mineralized in the new species). Although the osteology of B. pernix was not studied in detail (Pombal et al. 1998), on the basis of the available information its osteology is quite similar to that of the new species. See Figure 3 for osteological details of the new species. Body size (males SVL = 9.2���10.8 mm; females SVL = 11.1���12.4 mm) distinguishes the new species from Brachycephalus ferruginus (males SVL = 11.6���12.5 mm; females SVL = 13.0��� 14.5 mm; Alves et al. 2006), B. pernix (males SVL = 12.0��� 13.3 mm; females SVL = 14.1���15.8 mm; Pombal et al. 1998), and B. pombali (males SVL = 12.6���13.9 mm; females SVL= 14.6���15.3 mm; Alves et al. 2006). Also, in B. coloratus (males SVL = 10.3��� 10.6 mm; females SVL = 12.2���13.3 mm; Ribeiro et al. 2017), B. izecksohni (males SVL = 10.3���12.1 mm; females SVL = 12.5���13.1 mm; Ribeiro et al. 2005), and B. tridactylus (males SVL = 10.6���11.6; females SVL = 13.5���13.8 mm; Garey et al. 2012) males and/or females are slightly larger than in the new species. Brachycephalus actaeus sp. nov. is distinguishable from B. albolineatus, B. auroguttatus, B. boticario, B. fuscolineatus, B. leopardus, B. mariaeterezae, B. olivaceus, B. quiririensis, and B. verrucosus by a proportionally shorter head relative to body length (HL/SVL) and by a proportionally larger eye diameter related to head length (ED/HL). In B. actaeus sp. nov., HL/ SVL is 18���24% (x��= 20, SD = 1) and ED/HL is 52���73% (x��= 63, SD = 4), for the 32 adult specimens of the type series, without sexual distinction. In B. albolineatus, HL/SVL is 28���34% (x��= 31, SD = 4) and ED/HL is 36���42% (x��= 38, SD = 4); in B. auroguttatus, HL/ SVL is 29���38% (x��= 33, SD = 2) and ED/HL is 30���44% (x��= 33, SD = 3); in B. boticario, HL/ SVL is 31���36% (x��= 34, SD = 2) and ED/HL is 30���34% (x��= 32, SD = 1); in B. fuscolineatus, HL/ SVL is 29���34% (x��= 31, SD = 1) and ED/HL is 36���41% (x��= 39, SD = 1); in B. leopardus, HL/ SVL is 31���35% (x��= 33, SD = 1) and ED/HL is 34���43% (x��= 38, SD = 3); in B. mariaeterezae, HL/ SVL is 29���36% (x��= 33, SD = 2) and ED/HL is 36���42% (x��= 39, SD = 2); in B. olivaceus, HL/ SVL is 32���36% (x��= 34, SD = 1) and ED/HL is 26���36% (x��= 32, SD = 3); in B. quiririensis, HL/ SVL is 31���36% (x��= 34, SD = 1) and ED/HL is 28���34% (x��= 32, SD = 2); and in B. verrucosus, HL/ SVL is 30���36% (x��= 33, SD = 2) and ED/HL is 30���40% (x��= 34, SD = 3) (Pie & Ribeiro 2015; Ribeiro et al. 2015; Bornschein et al. 2016b). Additionally, Brachycephalus actaeus sp. nov. exhibits a rounded snout in dorsal and lateral views, which distinguishes it from B. brunneus, which has a slightly mucronate snout in dorsal view (Ribeiro et al. 2005); from B. leopardus, which has a slightly truncate snout in dorsal and lateral views (Ribeiro et al. 2015); and from B. quiririensis, which has a mucronate snout in dorsal view (Pie & Ribeiro 2015). Nostrils are not protuberant in B. auroguttatus, B. pernix, and B. fuscolineatus (Pombal et al. 1998; Ribeiro et al. 2015), which differ from the protuberant nostrils of the new species. In Brachycephalus tridactylus, finger IV is not externally visible (Garey et al. 2012), while in the new species it is reduced but externally distinct. In B. actaeus sp. nov., fingertips I, II, and IV are rounded, differing from B. pernix in which these fingertips are pointed (Pombal et al. 1998) and from B. brunneus, B. izecksohni, and B. leopardus, which have the tip of finger II pointed (Ribeiro et al. 2005; Ribeiro et al. 2015). The texture of the skin on the dorsum of Brachycephalus actaeus sp. nov. is rough; this characteristic distinguishes the new species from B. albolineatus, B. brunneus, B. coloratus, B. curupira, B. ferruginus, B. izecksohni, B. leopardus, B. pernix, B. pombali, and B. tridactylus, which have a smooth dorsum (Pombal et al. 1998; Ribeiro et al. 2005; Alves et al. 2006; Garey et al. 2012; Ribeiro et al. 2015; Bornschein et al. 2016b; Ribeiro et al. 2017). The dark green general color of Brachycephalus actaeus sp. nov. in life is very distinct from B. boticario, B. coloratus, B. ferruginus, B. fuscolineatus, B. izecksohni, B. leopardus, B. mariaeterezae, B. quiririensis, B. pernix, B. pombali, B. tridactylus, and B. verrucosus, which exhibit a bright yellow or orange general dorsal body color (Pombal et al. 1998; Ribeiro et al. 2005; Alves et al. 2006; Garey et al. 2012; Pie & Ribeiro 2015; Ribeiro et al. 2015; Ribeiro et al. 2017). The body color of the new species is similar to B. albolineatus, which has a greenish general coloration (Bornschein et al. 2016b), and B. olivaceus, which is predominantly dark green to brown (Ribeiro et al. 2015). The advertisement call of Brachycephalus actaeus sp. nov. differs in structure and frequency from all known advertisement calls within Brachycephalus. It is clearly distinct from B. crispus, B. darkside , B. ephippium, and B. pitanga by having higher frequencies together with a different call structure (Pombal et al. 1994; Ara��jo et al. 2012; Condez et al. 2014; Guimar��es et al. 2017). In these species, the advertisement call is characterized by the regular repetition of one low-frequency note with several pulses, while in B. actaeus sp. nov. the high-frequency notes are composed of just two pulses. In B. crispus, the notes generally last 0.28 seconds and are composed of 10 pulses; the frequency range is 3.5���5.7 kHz, while the dominant frequency is 4.6 kHz (Condez et al. 2014). In B. darkside, the average note duration is 0.11 seconds and notes are composed of six pulses; the frequency range is 2.5���5.8 kHz [considered dominant frequency in Guimar��es et al. (2017)], while the dominant frequency is 3.4 kHz [considered peak frequency in Guimar��es et al. (2017)]. In B. ephippium, notes typically last 0.12 seconds and are also composed of 12 pulses; the minimum and maximum frequencies are 3.4���5.3 kHz (Pombal et al. 1994). In B. pitanga, notes last 0.17 seconds and are composed of 11 pulses; the dominant frequency is 4.9 kHz (Ara��jo et al. 2012). The high frequency of the advertisement call of the new species is comparable to that described for B. hermogenesi and B. sulfuratus, the latter having the highest dominant frequency known for the genus (Condez et al. 2016). Nevertheless, the advertisement calls of these species differ from the new species in general structure, which is long and composed of a set of high-frequency notes. In B. hermogenesi, the call is composed of 1���5 notes; call lasts 0.2 seconds, with 1���5 pulses (Verdade et al. 2008). The dominant frequency in B. hermogenesi is 6.8 kHz (Verdade et al. 2008). In B. sulfuratus, the call is composed of 4���7 notes, each one lasting 0.19 seconds, with 9 pulses (Condez et al. 2016). The frequency range is 4.9���9.3 kHz and the dominant frequency is 6.7 kHz (Condez et al. 2016). When compared to the advertisement calls of its most closely related species, B. pernix and B. tridactylus, the new species call has shorter notes and higher frequencies. In B. pernix, notes last 0.03���0.06 seconds and are composed of three pulses; the frequency range is 4.5 kHz���6.7 kHz (Wistuba 1998). In B. tridactylus, the frequency range is 3.2���6.4 kHz and dominant frequency is 4.8 kHz (Garey et al. 2012). Description of holotype. Body robust, bufoniform; head wider than long; head length 19% of SVL; snout short, rounded in lateral and dorsal views (Figures 1A and 1B); nostrils protuberant; canthus rostralis indistinct; loreal region slightly concave; eyes slightly protruding laterally and dorsally; eye diameter 72% of head length; tympanum absent; lips nearly sigmoid; vocal sac not expanded externally; vocal slits present; tongue longer than wide, with the posterior half not adherent to floor of mouth; vomerine teeth absent; choanae small and ovoid, anterior to eyes. Arm and forearm moderately slender; hands with fingers I and IV reduced; finger II short but distinct; finger III large and robust; fingertips I, II, and IV rounded, fingertip III pointed; finger lengths IV Measurements of holotype (in mm). SVL 9.4; HL 1.8; HW 3.0; ND 0.4; IND 1.1; ED 1.3; IOD 2.1; END 0.6; AL 2.0; FAL 2.1; HAL 1.4; THL 3.8; TBL 3.3; FL 4.6. Color in life. (Figure 6) Iris black. General body color orange, dorsal surface of body covered by dark green blotches; in dorsal view, a poorly defined dark brown stripe extends from the interorbital region to the posterior end of the vertebral column; arms, legs, fingers III and IV and toe IV dark green, other fing, Published as part of Monteiro, Juliane Petry De Carli, Condez, Thais Helena, Garcia, Paulo Christiano De Anchietta, Comitti, Estev��o Jasper, Amaral, Ivan Borel & Haddad, C��lio Fernando Baptista, 2018, A new species of Brachycephalus (Anura, Brachycephalidae) from the coast of Santa Catarina State, southern Atlantic Forest, Brazil, pp. 483-505 in Zootaxa 4407 (4) on pages 486-500, DOI: 10.11646/zootaxa.4407.4.2, http://zenodo.org/record/1221060, {"references":["Izecksohn, E. (1971) Novo genero e nova especie de Brachycephalidae do estado do Rio de Janeiro, Brasil. Boletim do Museu Nacional, Zoologia, 280, 1 - 12.","Giaretta, A. A. & Sawaya, R. J. (1998) Second species of Psyllophryne (Anura: Brachycephalidae). Copeia, 1998, 985 - 987. https: // doi. org / 10.2307 / 1447345","Napoli, M. F., Caramaschi, U., Cruz, C. A. G. & Dias, I. R. (2011) A new species of flea-toad, genus Brachycephalus Fitzinger (Amphibia: Anura: Brachycephalidae), from the Atlantic rainforest of southern Bahia, Brazil. Zootaxa, 2739 (1), 33 - 40. https: // doi. org / 10.11646 / zootaxa. 2739.1.3","Condez, T. H., Monteiro, J. P. C., Comitti, E. J., Garcia, P. C. A., Amaral, I. B. & Haddad, C. F. B. (2016) A new species of flea-toad (Anura: Brachycephalidae) from southern Atlantic Forest, Brazil. Zootaxa, 4083 (1), 40 - 56. https: // doi. org / 10.11646 / zootaxa. 4083.1.2","Clemente-Carvalho, R. B. G., Antoniazzi, M. M., Jared, C., Haddad, C. F. B., Alves, A. C. R., Rocha, H. S., Pereira, G. R., Oliveira, D. F., Lopes, R. T. & Reis, S. F. (2009) Hyperossification in miniaturized toadlets of the genus Brachycephalus (Amphibia: Anura: Brachycephalidae): Microscopic structure and macroscopic patterns of variation. Journal of Morphology, 270 (11), 1285 - 1295. https: // doi. org / 10.1002 / jmor. 10755","Guimaraes, C. S., Luz, S., Rocha, P. C. & Feio, R. N. (2017) The dark side of pumpkin toadlet: a new species of Brachycephalus (Anura: Brachycephalidae) from Serra do Brigadeiro, southeastern Brazil. Zootaxa, 4258 (4), 327 - 344. https: // doi. org / 10.11646 / zootaxa. 4258.4.2","Haddad, C. F. B., Alves, A. C. R., Clemente-Carvalho, R. B. G. & Reis, S. F. (2010) A new species of Brachycephalus from the Atlantic Rain Forest in Sao Paulo state, southeastern Brazil (Amphibia: Anura: Brachycephalidae). Copeia, 2010, 410 - 420. https: // doi. org / 10.1643 / CH- 09 - 102","Pombal, J. P. (2010) A posicao taxonomica das \" variedades \" de Brachycephalus ephippium (Spix, 1824) descritas por Miranda- Ribeiro, 1920 (Amphibia, Anura, Brachycephalidae). Boletim do Museu Nacional, Zoologia, 526, 1 - 12.","Clemente-Carvalho, R. G. B., Klaczo, J., Perez, I., Alves, A. C. R., Haddad, C. F. B. & Reis, S. F. (2011) Molecular phylogenetic relationships and phenotypic diversity in miniaturized toadlets, genus Brachycephalus (Amphibia: Anura: Brachycephalidae). Molecular Phylogenetics and Evolution, 61, 79 - 89. https: // doi. org / 10.1016 / j. ympev. 2011.05.017","Condez, T. H., Clemente-Carvalho, R. B. G., Haddad, C. F. B. & Reis, S. F. (2014) A new species of Brachycephalus (Anura: Brachycephalidae) from the highlands of the Atlantic Forest, southeastern Brazil. Herpetologica, 70 (1), 89 - 99. https: // doi. org / 10.1655 / HERPETOLOGICA-D- 13 - 00044","Ribeiro, L. F., Alves, A. C. R., Haddad, C. F. B. & Reis, S. F. (2005) Two new species of Brachycephalus Gunther, 1985 from the state of Parana, southern Brazil (Amphibia, Anura, Brachycephalidae). Boletim do Museu Nacional, Zoologia, 519, 1 - 18.","Alves, A. C. R., Ribeiro, L. F., Haddad, C. F. B. & Reis, S. F. (2006) Two new species of Brachycephalus (Anura: Brachycephalidae) from the Atlantic Forest in Parana state, southern Brazil. Herpetologica, 62 (2), 221 - 233. https: // doi. org / 10.1655 / 05 - 41.1","Bornschein, M. R., Ribeiro, L. F., Blackburn, D. C., Stanley, E. L. & Pie, M. R. (2016 b). A new species of Brachycephalus (Anura: Brachycephalidae) from Santa Catarina, southern Brazil. PeerJ, 4 (e 2629), 1 - 19. https: // doi. org / 10.7717 / peerj. 2629","Ribeiro, L. F., Blackburn, D. C., Stanley, E. L., Pie, M. R. & Bornschein M. R. (2017) Two new species of the Brachycephalus pernix group (Anura: Brachycephalidae) from the state of Parana, southern Brazil. PeerJ, 5 (e 3603), 1 - 28. https: // doi. org / 10.7717 / peerj. 3603","Pombal, J. P., Wistuba, E. M. & Bornschein, M. R. (1998) A new species of brachycephalid (Anura) from the Atlantic Rain Forest of Brazil. Journal of Herpetology, 32 (1), 70 - 74. https: // doi. org / 10.2307 / 1565481","Garey, M. V., Lima, A. M. X., Hartmann, M. T. & Haddad, C. F. B. (2012) A new species of miniaturized toadlet, genus Brachycephalus (Anura: Brachycephalidae), from southern Brazil. Herpetologica, 68 (2), 266 - 271. https: // doi. org / 10.1655 / HERPETOLOGICA-D- 11 - 00074.1.","Araujo, C. B., Guerra, T. J., Amatuzzi, M. C. O. & Campos, L. A. (2012) Advertisement and territorial calls of Brachycephalus pitanga (Anura: Brachycephalidae). Zootaxa, 3302, 66 - 67.","Verdade, V. K., Rodrigues, M. T., Cassimiro, J., Pavan, D., Liou, N. & Lange, M. (2008) Advertisement call, vocal activity, and geographic distribution of Brachycephalus hermogenesi (Giaretta and Sawaya, 1998) (Anura, Brachycephalidae). Journal of Herpetology, 42 (3), 542 - 549. https: // doi. org / 10.1670 / 07 - 287.1","Goutte, S., Mason, M. J., Christensen-Dalsgaard, J., Montealegre-Z, F., Chivers, B. D., Sarria-S, F. A., Antoniazzi, M. M., Jared, C., Sato, L. A. & Toledo, L. F. (2017) Evidence of auditory insensitivity to vocalization frequencies in two frogs. Scientific Reports, 7, 12121. https: // doi. org / 10.1038 / s 41598 - 017 - 12145 - 5","Pie, M. R., Meyer, A. L. S., Firkowski, C. R., Ribeiro, L. F. & Bornschein, M. R. (2013) Understanding the mechanisms underlying the distribution of microendemic montane frogs (Brachycephalus spp., Terrarana: Brachycephalidae) in the Brazilian Atlantic Rainforest. Ecological Modelling, 250, 165 - 176. https: // doi. org / 10.1016 / j. ecolmodel. 2012.10.019","Bornschein, M. R., Firkowski, C. R., Belmonte-Lopes, R., Correa, L., Ribeiro, L. F., Morato, S. A. A., Antoniazzi-Junior, R. L., Reinert, B. L., Meyer, A. L. S., Cini, F. A. & Pie, M. R. (2016 a). Geographical and altitudinal distribution of Brachycephalus (Anura: Brachycephalidae) endemic to the Brazilian Atlantic Rainforest. PeerJ, 4 (e 2490), 1 - 41. https: // doi. org / 10.7717 / peerj. 2490","Oliveira, J. C. F., Coco, L., Pagotto, R. V., Pralon, E., Vrcibradic, D., Pombal, J. P. & Rocha, C. F. D. (2012) Amphibia, Anura, Brachycephalus didactylus (Izecksohn, 1971) and Zachaenus parvulus (Girard, 1853): Distribution extension. Check List, 8 (2), 242 - 244. https: // doi. org / 10.15560 / 8.2.242","Clemente-Carvalho, R. B. G., Monteiro, L., Bonato, V., Rocha, H., Pereira, G., Oliveira, D., Lopes, R., Haddad, C. F. B., Martins, E. & Reis, S. F. (2008) Geographic variation in cranial shape in the pumpkin toadlet (Brachycephalus ephippium): a geometric analysis. Journal of Herpetology, 42 (1), 176 - 185. https: // doi. org / 10.1670 / 07 - 141 R 1.1","Pimenta, B. V. S., Bernils, R. S. & Pombal, J. P. (2007) Amphibia, Anura, Brachycephalidae, Brachycephalus hermogenesi: filling gap and geographic distribution map. Check List, 3 (3), 277 - 279. https: // doi. org / 10.15560 / 3.3.277","Pombal, J. P. & Izecksohn, E. (2011) Uma nova especie de Brachycephalus (Anura: Brachycephalidae) do estado do Rio de Janeiro. Papeis Avulsos de Zoologia, 51 (28), 443 - 451. https: // doi. org / 10.1590 / S 0031 - 10492011002800001","Clemente-Carvalho, R. B. G., Perez, S. I., Tonhati, C. H., Condez, T. H., Sawaya, R. J., Haddad, C. F. B. & Reis, S. F. (2015) Boundaries of morphological and molecular variation and the distribution range of a miniaturized froglet, Brachycephalus nodoterga (Anura: Brachycephalidae). Journal of Herpetology, 50 (1), 169 - 178. https: // doi. org / 10.1670 / 14 - 119","Abegg, A. D., Ortiz, F. R., Rocha, B. & Condez, T. H. (2015) A new record for Brachycephalus nodoterga (Amphibia, Anura, Brachycephalidae) in the state of Sao Paulo. Brazil. Check List, 11 (1769), 1 - 3. https: // doi. org / 10.15560 / 11.5.1769"]}

We describe a new species of Brachycephalus from municipality of São Francisco do Sul and municipality of Itapoá, in the state of Santa Catarina, Brazil, southern Atlantic Forest. The new species is known from six localities from near sea level up to 250 meters and represents the first record of a “pumpkin-toadlet” occurring in the lowlands. Morphological traits and phylogenetic analysis of a fragment the 16S mtDNA gene place the new species in the Brachycephalus pernix group. The new species is supported by external morphology, osteology, advertisement call, and mtDNA divergence. It is characterized, among other traits by a dorsal body color dark green with a dark brown vertebral stripe, and an orange background; snout-vent length of 9.2–10.8 mm in males and 11.1–12.4 mm in females; and advertisement call short (0.02–0.03 seconds), composed of one high-frequency note (dominant frequency 6.6–7.3 kHz). We observed synchronized alternation in the emission of vocalizations among neighbor males, indicating that males of the new species are able to hear and use vocalizations to interact with each other. We provide descriptions of clutch, eggs, and juvenile and observations on parental care. The new species has not been recorded within any protected area and can be threatened by human-induced habitat loss and modification.

The amphibian fauna of the state of Rio de Janeiro, in southeatern Brazil, is characterized by high species richness and rates of endemism, and is still insufficiently known. A first list of amphibian species with occurrence in the state was published in 2004 and reported 166 taxa, but since then many new records, descriptions of new taxa, and revalidations and synonymizations of species have consistently improved the knowledge about the state’s amphibian biodiversity. Thus, a review and update of that list was deemed necessary. We herein present an updated and commented list of amphibian species occurring in the state of Rio de Janeiro based on a survey of the literature. We recorded the occurrence of a total of 201 species of amphibians (197 anurans and four caecilians) in Rio de Janeiro, with 54 of them (ca. 27%) considered to be endemic of the state. Our study presents an increase in species richness of 21% since the publication of the previous list, indicating a consistent advance in knowledge of the composition of the amphibian fauna in the state. In spite of its relatively small territorial extension (total area ca. 43,800 km²), the state of Rio de Janeiro contains nearly 20% of the amphibian species known to occur in Brazil and around 40% of those occurring in the Atlantic Forest biome. Thus, that state constitutes an important reservoir of amphibian biodiversity in the Atlantic Forest biome and in Brazil, as a whole.